Fungicide Resistance Profiles of Botrytis cinerea Isolates From Michigan Vineyards and Development of a TaqMan Assay for Detection of Fenhexamid Resistance.

Botrytis cinerea on grapes causes bunch rot at both pre- and postharvest stages, in which losses can reach up to 100%. Chemical control primarily relies on the prophylactic use of site-specific fungicides. Repeated applications of these products raise the risk of fungicide resistance development in B. cinerea populations, which can result in disease control failures. To determine the extent of resistance, B. cinerea isolates were collected from grape clusters in the northwest and southwest grape growing regions of Michigan in 2014 and 2018 (n = 115 and 125, respectively). These isolates were phenotyped using discriminatory doses of eight fungicides to determine the levels of resistance. Fungicide resistance increased from 2014 to 2018, mostly affecting the active ingredients fenhexamid, fluopyram, and iprodione. B. cinerea isolates resistant to multiple fungicides were detected in 2014 and 2018, with a higher frequency of resistance in 2018. TaqMan real-time PCR has been developed to detect B. cinerea fungicide resistance to fenhexamid and to differentiate the erg27 F412S/I/V alleles. The TaqMan assay was tested for sensitivity, specificity, and reproducibility on purified DNA and infected grape tissue samples. Our data provide essential information to growers about the efficacy for B. cinerea control using the available botryticides. Furthermore, the developed fenhexamid markers will be transferred to diagnostic clinics to assist growers in the management of bunch rot before resistance-related control failures occur.

Ethical Use and Impact of Participatory Approaches to Research in Post-Disaster Environments: An Australian Bushfire Case Study.

This paper presents a case study of Beyond Bushfires, a large, multisite, mixed method study of the psychosocial impacts of major bushfires in Victoria, Australia. A participatory approach was employed throughout the study which was led by a team of academic investigators in partnership with service providers and government representatives and used on-site visits and multiple methods of communication with communities across the state to inform decision-making throughout the study. The ethics and impacts of conducting and adapting the approach within a post-disaster context will be discussed in reference to theories and models of participatory health research. The challenges of balancing local interests with state-wide implications will also be explored in the description of the methods of engagement and the study processes and outcomes. Beyond Bushfires demonstrates the feasibility of incorporating participatory methods in large, post-disaster research studies and achieving rigorous findings and multilevel impacts, while recognising the potential for some of the empowering aspects of the participatory experience to be reduced by the scaled-up approach.

Hyperbaric oxygen for neurologic indications--action plan for multicenter trials in: stroke, traumatic brain injury, radiation encephalopathy & status migrainosus.

The 2008 Toronto Hyperbaric Medicine Symposium was convened to discuss research into neurologic indications for hyperbaric oxygen therapy (HBO2T). Four topics were particularly addressed: acute ischemic stroke; acute traumatic brain injury; brain radiation necrosis; and status migrainosus. Four multicenter trials were designed and proposed to evaluate the efficacy of HBO2T for these indications and are presented here in addition to brief reviews of the rationale behind each.

Regulation of Junonia coenia densovirus P9 promoter expression.

Transcriptional activity of the Junonia coenia densovirus (JcDNV) P9 promoter depends on a 557-bp sequence located within the overlapping 3' sequences for viral capsid and nonstructural genes. Utilizing a somatic transformation assay to assess JcDNV promoter activity in Drosophila melanogaster and Plodia interpunctella, viral sequences were subjected to deletional analysis. Removal of a 685-bp fragment reduced P9-driven expression to background levels. Inclusion of a second expression cassette demonstrated vector persistence and confirmed somatic transformation. P9 promoter-driven expression was restored by insertion of a 557-bp JcDNV fragment or by inclusion of a heterologous baculovirus hr5 enhancer. Consensus polycomb transcriptional factor binding sites were identified within the 557-bp fragment, which suggests a potential role in regulating densoviral transcription.

Somatic transformation efficiencies and expression patterns using the JcDNV and piggyBac transposon gene vectors in insects.

A somatic transformation gene vector that exploits the genomic integration properties of Junonia coenia lepidopteran densovirus (JcDNV) sequences in vivo has been developed. JcDNV somatic transformation vectors are derivatives of plasmids containing an interrupted genome of JcDNV that provide efficient, robust vectors that can be used to examine regulation of chromosomally integrated transgenes in insects. Microinjection of JcDNV plasmids into syncytial embryos of Drosophila melanogaster or the lepidopterans Plodia interpunctella, Ephestia kuehniella or Trichoplusia ni resulted in persistent transgene expression throughout development. Inclusion of transgenes with tissue-specific promoters resulted in expression patterns canonical with phenotypes of piggyBac germline transformants. Somatic transformation required the presence of the viral inverted terminal repeat in cis only and did not depend upon non-structural viral proteins.

The relationship between HIV seroconversion illness, HIV test interval and time to AIDS in a seroconverter cohort.

Seroconversion illness is known to be associated with more rapid HIV disease progression. However, symptoms are often subjective and prone to recall bias. We describe symptoms reported as seroconversion illness and examine the relationship between illness, HIV test interval (time between antibody-negative and anibody-positive test dates) and the effect of both on time to AIDS from seroconversion. We used a Cox model, adjusting for age, sex, exposure group and year of estimated seroconversion. Of 1820 individuals, information on seroconversion illness was available for 1244 of whom 423 (34%) reported symptomatic seroconversion. Persons with a short test interval (< or = 2 months) were significantly more likely to report an illness than people with a longer interval (OR 6.76, 95% CI 4.75-9.62). Time to AIDS was significantly faster (P = 0.01) in those with a short test interval. The HIV test interval is a useful replacement for information on seroconversion illness in studies of HIV disease progression.

First Report of Tobacco ringspot virus in Table Grapes in Michigan.

In 1998, several 10-year-old 'Marquis' and 'Vanessa' (Vitis sp.) table grapevines at the Southwest Michigan Research and Extension Center in Benton Harbor started showing decline symptoms such as stunted shoots with small leaves and berries. Vines eventually stopped producing fruit and died. In 1999, symptomatic vines were indexed on cucumber (Cucumis sativus L. 'National Pickling') cotyledons, which developed chlorotic local lesions. Symptomatic cucumber tissue was tested using Ouchterlony double diffusion with polyclonal antibodies for Tomato ringspot virus (ToRSV) and Tobacco ringspot virus (TRSV) provided by D. Ramsdell. Samples tested positive for TRSV. 'Marquis' vines (9 of 45) in the affected area also tested positive in double-antibody sandwich enzyme-linked immunosorbent assay with polyclonal antibodies for TRSV (Agdia, Inc., Elkhart, IN). Analysis of soil samples from the site in 1989 yielded five dagger nematodes (Xiphinema americanum Cobb) per g of soil, confirming the potential for virus spread by nematodes (1). TRSV reportedly infects Vitis vinifera L. but not V. labrusca L. (1). 'Marquis' and 'Vanessa' have V. vinifera heritage. The detection of TRSV has led to the establishment of a program for the production of virus-tested table grape planting stock, as well as research on the utility of nematode-resistant rootstocks for growing table grapes at the infested site. Reference: (1) R. C. Pearson and A. C. Goheen, eds. Compendium of Grape Diseases, The American Phytopathological Society, St. Paul, MN, 1995.

Cloning, sequencing, and promoter identification of Blueberry red ringspot virus, a member of the family Caulimoviridae with similarities to the "Soybean chlorotic mottle-like" genus.

The double-stranded DNA genome of Blueberry red ringspot virus (BRRV), a member of the family Caulimoviridae, was cloned and sequenced. The genome organization and relationships of the 8303 nt sequence revealed BRRV to be a tentative member of the genus that has been provisionally named "Soybean chlorotic mottle-like viruses", rather than a member of the genus Caulimovirus, in which it had been placed previously. Insertion of the putative 35S promoter homolog of BRRV into promoterless constructs carrying the UidA (beta-glucuronidase) gene resulted in high-level transient expression from cranberry and stable expression from transgenic tobacco. Sequences of 5'-RACE clones derived from transcripts from transgenic tobacco were consistent with the map position of the promoter.

Capitation and primary care in Canada: financial incentives and the evolution of health service organizations.

Alternative approaches to the funding, organization, and delivery of primary care have been the subject of ongoing discussion and debate in many industrialized nations for many years. One common recommendation has been to use capitation, as opposed to fee-for-service, as the payment method for physicians. In this study the authors use data from interviews with physicians and Ministry of Health officials to trace the evolution of Ontario's Health Service Organization (HSO) program, the only program of capitation-funded physician care in Canada. The program has developed in three phases: formation in the early 1970s, expansion in the late 1970s and throughout the 1980s, and restructuring in the 1990s. The analysis focuses on the perceptions and actions of policymakers and physicians who became involved with the program at different points in its evolution, and identifies how they perceived and responded to the financial incentives that were introduced to promote the program. This case study allows an examination of the shifting objectives, communications, perceptions, and responses of policymakers and stakeholders in changing contexts over a period of more than 20 years. The long history of the HSO program provides the opportunity to examine the factors that can cause financial incentives to go awry. The authors suggest how this case study offers lessons for financial incentive policymaking.

Cerebral sinovenous thrombosis in children.

BACKGROUND: Cerebral sinovenous thrombosis in children is a serious disorder, and information is needed about its prevention and treatment. METHODS: The Canadian Pediatric Ischemic Stroke Registry was initiated in 1992 at the 16 pediatric tertiary care centers in Canada. Children (newborn to 18 years of age) with symptoms and radiographic confirmation of sinovenous thrombosis were included. RESULTS: During the first six years of the registry, 160 consecutive children with sinovenous thrombosis were enrolled, and the incidence of the disorder was 0.67 cases per 100,000 children per year. Neonates were most commonly affected. Fifty-eight percent of the children had seizures, 76 percent had diffuse neurologic signs, and 42 percent had focal neurologic signs. Risk factors included head and neck disorders (in 29 percent), acute systemic illnesses (in 54 percent), chronic systemic diseases (in 36 percent), and prothrombotic states (in 41 percent). Venous infarcts occurred in 41 percent of the children. Fifty-three percent of the children received antithrombotic agents. Neurologic deficits were present in 38 percent of the children, and 8 percent died; half the deaths were due to sinovenous thrombosis. Predictors of adverse neurologic outcomes were seizures at presentation and venous infarcts. CONCLUSIONS: Sinovenous thrombosis in children affects primarily neonates and results in neurologic impairment or death in approximately half the cases. The occurrence of venous infarcts or seizures portends a poor outcome.

Lay constructions of HIV and complementary therapy use.

This study examines the meanings that individuals with HIV attach to their use of complementary therapies. A qualitative analysis of 66 interviews completed between 1993 and 1998 showed that complementary therapies represent different things for these individuals--a health maintenance strategy, a healing strategy, an alternative to Western medicine, a way of mitigating the side-effects of drug therapies, a strategy for maximizing quality of life, a coping strategy, and a form of political resistance. We found that the meanings individuals ascribe to complementary therapies and the benefits they expect to derive from them are not idiosyncratic, but linked to social characteristics--sexuality, ethnocultural background, gender--and to beliefs about health and illness, values and experiences. We found as well that these meanings are neither mutually exclusive nor fixed. The therapies often appeal to individuals on different levels and their appeal may change over time.

Obstetrics in general practice. A rural perspective.

Obstetrics is one of the most demanding yet rewarding disciplines required of rural general practice although rising medical indemnity fees combined with medico-legal interest in this discipline make the current rural generalist obstetricians a threatened species. These factors, plus the difficulty generalist registrars have in obtaining good procedural training in this specialist field make obstetric practice less than attractive for future rural general practitioners. This article provides a brief overview of rural obstetric practice from the perspective of a rural generalist obstetrician with 18 years experience. Case studies are used to highlight the challenges of this field.

Isolated sulfite oxidase deficiency.

Isolated sulfite oxidase (SO) deficiency is an autosomal recessively inherited inborn error of sulfur metabolism. In this report of a ninth patient the clinical history, laboratory results, neuropathological findings and a mutation in the sulfite oxidase gene are described. The data from this patient and previously published patients with isolated sulfite oxidase deficiency and molybdenum cofactor deficiency are summarized to characterize this rare disorder. The patient presented neonatally with intractable seizures and did not progress developmentally beyond the neonatal stage. Dislocated lenses were apparent at 2 months. There was increased urine excretion of sulfite and S-sulfocysteine and a decreased concentration of plasma cystine. A lactic acidemia was present for 6 months. Liver sulfite oxidase activity was not detectable but xanthine dehydrogenase activity was normal. The boy died of respiratory failure at 32 months. Neuropathological findings of cortical necrosis and extensive cavitating leukoencephalopathy were reminiscent of those seen in severe perinatal asphyxia suggesting an etiology of energy deficiency. A point mutation that resulted in a truncated protein missing the molybdenum-binding site has been identified.

Declaring pediatric brain death: current practice in a Canadian pediatric critical care unit.

OBJECTIVE: To document the criteria used to declare brain death in a pediatric critical care unit (PCCU). DESIGN: Retrospective chart review. SETTING: Regional PCCU in southwestern Ontario. PATIENTS: Sixty patients 16 years of age or less declared brain dead from January 1987 through December 1992. OUTCOME MEASURES: Presence or absence of documentation of irreversible deep coma, nonresponsive cranial nerves, absent brain-stem reflexes, persistent apnea after removal from ventilator, presence or absence of blood flow detected by radioisotope scanning, presence or absence of electroencephalographic evidence of electrocerebral activity. RESULTS: The 60 patients accounted for 1.5% of all PCCU admissions; 17 were under 1 year of age. In 39 cases brain death was diagnosed using clinical criteria ("certified brain death"), which could not be fully applied in the remaining 21 cases ("uncertifiable but suspected brain death"). Electroencephalography and cerebral blood-flow studies with technetium-99m hexamethyl-propyleneamine oxime were used as ancillary tests in 16 patients with certified brain death and in 17 with uncertifiable but suspected brain death who survived long enough to be tested. Electrocerebral silence was demonstrated in all nine patients who underwent electroencephalography. Cerebral blood flow was undetectable in 26 of the 30 patients tested, and an abnormal pattern of blood flow was seen in the remaining 4, all of whom received a diagnosis of certified brain death. CONCLUSIONS: Pediatricians in this large tertiary care referral centre are using clinical criteria based on the 1987 guidelines of the CMA to diagnose brain death in pediatric patients, including neonates. When clinical criteria cannot be fully applied, ancillary methods of investigation are consistently used. Although the soundness of this pattern of practice is established for adults and older children, its applicability to neonates and infants still needs to be validated.

Nerve biopsy findings in Niemann-Pick type II (NPC)

The severe infantile form of Niemann-Pick disease type II was diagnosed in a 4-year-old girl and confirmed by demonstrating in cultured skin fibroblasts a deficiency of low-density lipoprotein-stimulated cholesterol ester synthesis of < 5% of normal. Electrodiagnostic studies revealed changes of a predominantly demyelinating motor and sensory polyneuropathy. Light microscope and ultrastructural examination of a peroneal nerve biopsy showed unique changes. Compacted myelin sheaths were disproportionately thin with marked globular irregularities in single teased nerve fibres and evidence of chronic demyelination. The majority of axons were preserved but axonal spheroids and cytoskeletal abnormalities akin to neuroaxonal dystrophy were noted. Membrane-bound multi-lobulated lysosomal inclusions of floccular and electron-dense material were present in Schwann cells (SC), endoneurial fibroblasts, macrophages, pericytes and endothelial cells. SC of myelinated fibres were stuffed with whorls of concentric osmiophilic membraneous profiles and electron-lucent material. The findings are diagnostic and differ from those of classical Niemann-Pick disease.

Comparison of alternative models for predicting the uptake of chlorinated hydrocarbons by oligochaetes.

The uptake of chlorinated hydrocarbons from contaminated sediments by oligochaetes was analyzed using three alternative models. Uptake appears to be best explained using a model that explicates the relationship of oligochaete body burden as a function of weight-dependent feeding rates, fractional absorption, and exposure times. The model explains observed variations in body burdens with oligochaete weights. Oligochaetes appear to absorb small fractions of ingested hydrocarbons. Fractional absorption of ingested contaminants appears to have a parabolic relationship with octanol-water partition coefficients and may decrease over time.

Uncertainty in risk assessment: exceedence frequencies, acceptable risk, and risk-based decision making.

The use of "exceedence frequencies" is proposed as a means of evaluating the acceptability of uncertain environmental risks. A survey was conducted of U.S. Environmental Protection Agency personnel. When respondents were requested to identify preferred "acceptable" exceedence frequencies at four different regulatory risk thresholds (10(-3) to 10(-6), three general response patterns were found: "absolutist" responses in which no level of exceedence was deemed acceptable; "fixed" responses in which respondents felt that the level of exceedence should remain constant at all risk levels; and "outcome sensitive" responses in which preferred exceedence frequencies were inversely related to the risk level. This last group's responses were based on the rationale that the "marginal cost" of risk (in terms of expected mortalities) increases as the risk threshold increases. Although the majority of respondents accepted the concept of an acceptable risk exceedence frequency, the low exceedence frequencies selected as being "acceptable" by survey respondents (generally 5% or less) appear to be overly conservative given the degree of uncertainty inherent in risk assessments. Finally, two regulatory uses of exceedence frequencies are proposed as a tool for streamlined risk ranking.