Development of Weather-Based Predictive Models for Fusarium Head Blight and Deoxynivalenol Accumulation for Spring Malting Barley.

The associations between Fusarium head blight (FHB), caused by Gibberella zeae, and deoxynivalenol (DON) accumulation in spring malting barley (Hordeum vulgare) and hourly weather conditions predictive of DON accumulation were examined using data from six growing seasons in the U.S. Northern Great Plains. Three commonly grown cultivars were planted throughout the region, and FHB disease and DON concentration were recorded. Nine predictor variables were calculated using hourly temperature and relative humidity during the 10 days preceding full head spike emergence. Simple logistic regression models were developed using these predictor variables based on a binary threshold for DON of 0.5 mg/kg. Four of the nine models had sensitivity greater than 80%, and specificity of these models ranged from 67 to 84% (n = 150). The most useful predictor was the joint effect of average hourly temperature and a weighted duration of uninterrupted hours (h) with relative humidity greater than or equal to 90%. The results of this study confirm that FHB incidence is significantly associated with DON accumulation in the grain and that weather conditions prior to full head emergence could be used to accurately predict the risk of economically significant DON accumulation for spring malting barley.

Widespread Occurrence of the Aecial Stage of Sunflower Rust Caused by Puccinia helianthi in North Dakota and Minnesota in 2008.

Sunflower rust caused by Puccinia helianthi (Schw.) is widespread in North America and occurs annually on cultivated sunflower (Helianthus annuus L.) and wild annual and perennial Helianthus spp., although severity on the U.S. sunflower crop has been increasing in recent years (2). P. helianthi is a autoecious, macrocyclic rust, but the aecial stage is rarely observed in the field (1,3,4). In most years, the earliest appearance of sunflower rust in North Dakota (ND) and surrounding states usually occurs in early August as the uredinial stage. Initial inoculum can result from urediniospores that overwinter in the Northern Great Plains, urediniospores blown in from areas south of North Dakota, or basidiospores completing the life cycle. However, aecia have been noted very infrequently and never widespread, indicating initial inoculum is usually urediniospores. Aecia of P. helianthi were first observed on 24 June 2008 in a commercial sunflower field (confection hybrid CHS 3126) near Mohall, ND. Aecia cups measuring 0.2 to 0.3 mm in diameter were observed in clusters that were 1 to 7 mm wide in diameter and containing as many as 150 cups. Aecia cups were bright orange but turned brown-black as they senesced. As many as 15 aecial clusters occurred on individual leaves or cotyledons. Aeciospores were ellipsoid, hyaline, and measured approximately 20 µm in diameter. On 4 July 2008, uredinia were first observed in the same Mohall, ND field. At that time, uredinia, aecia, and senesced aecia could all be found on the same leaves. In a non-fungicide-treated strip of the field, pustule coverage on the lower leaves was 10 to 20% by mid-July, pustule coverage on the upper four leaves of plants in the untreated strip was 56% by 27 August, and yield at harvest was less than 200 kg/ha. The rest of the field was sprayed twice with fungicides and yielded 1,571 kg/ha, which is similar to the statewide yield average of 1,573 kg/ha in 2008. To determine the prevalence of aecia in the primary growing regions of ND and Minnesota (MN), surveys were conducted in 75 sunflower fields in 18 counties between 22 and 24 July in ND and 34 fields in 8 counties between 17 and 31 July in MN. Incidence of aecia and uredinia were determined by visual observation of a minimum of 20 plants scouted in a 'W' pattern in the field. Rust was found in 31 and 53% of fields in ND and MN, respectively. In fields where rust was found, both aecia and uredinia were observed in 37% of the fields, aecia only in 29% of the fields, and uredinia only in 34% of the fields. Although it is uncertain why aecia were widespread in 2008, night temperatures in Mohall, ND, where aecia were first observed, reached the dew point temperature on 51 of 61 days in June and July, suggesting that dew or fog likely formed. Thus, favorable conditions for germination and infection early in the growing season indicate widespread occurrence of rust was likely a result of local inoculum sources. The early appearance of aecia is cause for concern for two reasons: significant yield loss can occur when rust appears early in the growing season; and the presence of aecia suggest that the pathogen completed its sexual cycle. When P. helianthi completes its life cycle it is likely that a greater diversity of races will result (4). References: (1) D. L. Bailey. Univ. Minn. Tech. Bull. 16:1, 1923. (2) D. Berglund. Natl. Sunflower Assoc. Online publication. /Berglund_2007_NSA_Survey_08. 2008. (3) H. S. Jackson. Mem. Torrey Bot. Club 18:1, 1931. (4) G. A. Kong et al. Australas. Plant Pathol. 28:320, 1999.

Efficacy of Oral Encochleated Amphotericin B in a Mouse Model of Cryptococcal Meningoencephalitis.

Cryptococcus neoformans is an encapsulated yeast responsible for approximately a quarter of a million deaths worldwide annually despite therapy, and upwards of 11% of HIV/AIDS-related deaths, rivaling the impact of tuberculosis and malaria. However, the most effective antifungal agent, amphotericin B, requires intravenous delivery and has significant renal and hematopoietic toxicity, making it difficult to utilize, especially in resource-limited settings. The present studies describe a new nanoparticle crystal encapsulated formulation of amphotericin B known as encochleated amphotericin B (CAmB) that seeks to provide an oral formulation that is low in toxicity and cost. Using a 3-day delayed model of murine cryptococcal meningoencephalitis and a large inoculum of a highly virulent strain of serotype A C. neoformans, CAmB, in combination with flucytosine, was found to have efficacy equivalent to parental amphotericin B deoxycholate with flucytosine and superior to oral fluconazole without untoward toxicity. Transport of fluorescent CAmB particles to brain as well as significant brain levels of amphotericin drug was demonstrated in treated mice, and immunological profiles were similar to those of mice treated with conventional amphotericin B. Additional toxicity studies using a standardized rat model showed negligible toxicity after a 28-day treatment schedule. These studies thus offer the potential for an efficacious oral formulation of a known fungicidal drug against intrathecal cryptococcal disease.IMPORTANCECryptococcus neoformans is a significant global fungal pathogen that kills an estimated quarter of a million HIV-infected individuals yearly and has poor outcomes despite therapy. The most effective therapy, amphotericin B, is highly effective in killing the fungus but is available only in highly toxic, intravenous formulations that are unavailable in most of the developing world, where cryptococcal disease in most prevalent. For example, in Ethiopia, reliance on the orally available antifungal fluconazole results in high mortality, even when initiated as preemptive therapy at the time of HIV diagnosis. Thus, alternative agents could result in significant saving of lives. Toward this end, the present work describes the development of a new formulation of amphotericin B (CAmB) that encapsulates the drug as a crystal lipid nanoparticle that facilitates oral absorption and prevents toxicity. Successful oral absorption of the drug was demonstrated in a mouse model that, in combination with the antifungal flucytosine, provided efficacy equal to a parental preparation of amphotericin B plus flucytosine. These studies demonstrate the potential for CAmB in combination with flucytosine to provide an effective oral formulation of a well-known, potent fungicidal drug combination.

Agronomic and Economic Responses of Spring Wheat to Management of Fusarium Head Blight.

Spring wheat (Triticum aestivum) crop losses in the Red River Valley of Minnesota and North Dakota caused by Fusarium head blight (FHB) epidemics incited by Fusarium graminearum are common. Fungicide application is often recommended when environments promote disease development but benefits have not been fully evaluated when environment, cultivar resistance, and economic outcome are considered. Agronomic and economic characters were determined for cultivars with various resistance levels when treated with no fungicide; propiconazole at 63 g active ingredient (a.i.)/ha applied at Feekes growth stage (FGS) 2, tebuconazole at 126 g a.i./ha applied at FGS 10.51, or propiconazole at 63 g a.i./ha applied at FGS 2 followed by tebuconazole at 126 g a.i./ha applied at FGS 10.51. Revenue returned from FHB moderately susceptible (MS) cultivars was 8% greater than moderately resistant (MR) cultivars in low-disease environs but differences were not significant when disease was moderate. Deoxynivalenol accumulation in grain of MS and MR cultivars was unchanged by fungicide treatment. MS cultivars were economically more adventitious to grow than MR cultivars in both disease environments.

Region and Field Level Distributions of Aster Yellows Phytoplasma in Small Grain Crops.

Aster yellows (AY), a disease of small grain crops caused by aster yellows phytoplasma (AYp), produces disease symptoms similar to barley yellow dwarf (BYD). From 2003 to 2005, small grain production fields in Minnesota and North Dakota were surveyed to determine the incidences of AY and BYD. In-field spatial patterns of AY-infected plants also were investigated. Plants collected along a five-point transect line were tested for AYp using nested polymerase chain reaction (PCR) and quantitative real-time PCR assays, and extracted plant sap was tested for serotypes PAV and RPV of Barley yellow dwarf virus (BYDV) using enzyme-linked immunosorbent assays. During 2003, 2004, and 2005, AYp was detected in plants from 49, 15, and 7% of tested fields, respectively, whereas BYDV was found in plants from 2, 0, and 5% of fields, respectively. Average amplicon count number indicated an in-field spatial trend for greater incidence of AYp and increased populations of AYp in plants located near field edges, with comparably low copy numbers at transect point locations toward the direction of field center. AY is likely a common but largely undetected disease on small grain crops in the Upper Midwest.

Distribution of Phoma sclerotioides on Alfalfa and Winter Wheat Crops in the North Central United States.

Brown root rot of alfalfa (Medicago sativa), caused by Phoma sclerotioides, has been reported in several states in the northern United States and in western Canada. A survey was conducted to determine the distribution of the fungus in Minnesota and Wisconsin. Isolates of the pathogen were recovered from roots of alfalfa, winter wheat, and perennial ryegrass plants. The internal transcribed spacer (ITS) 1, 5.8S, and ITS2 of the rDNA of the isolates from alfalfa and wheat were identical and matched the sequences of a P. sclerotioides isolate from Wyoming. The fungus was found to be widespread in both states and was detected in roots of alfalfa plants from 17 counties in Minnesota and 14 counties in Wisconsin using polymerase chain reaction (PCR)-based assays. A real-time PCR assay was developed that increased sensitivity of detecting the pathogen from plant tissues and soil. The isolates from alfalfa caused disease on inoculated winter wheat plants. Although the fungus was previously found associated with roots of diseased cereal and turfgrass plants, this is the first demonstration of pathogenicity of P. sclerotioides on wheat.

Patient progress rounds on a university hospital cardiology service.

The authors describe patient progress rounds, a weekly meeting of representatives of all disciplines associated with a university hospital cardiology service. The objective is coordination of treatment for all patients in the interest of providing comprehensive care. Each patient's clinical status is reviewed, but primary attention is given to psychosocial aspects. These rounds provide a medium for identifying and dealing with patients' needs, a cooperative teaching and learning venture for staff, a multidisciplinary team approach that increases quality of care, and an opportunity for doctors to confront the reality of their patients as psychosocial beings. Case vignettes illustrate the use and effectiveness of patient progress rounds.

Human T-cell lymphotropic virus infection among blood donors in south Florida. The Transfusion Safety Study Group.

Knowledge of the epidemiologic pattern of human T-lymphotropic virus (HTLV) in the United States is being enlarged by blood donor screening. We tested stored sera from 29,937 donations made in South Florida in 1984-1985. Twenty-three donors were confirmed as seropositive, a prevalence of 0.8 per 1,000 donations. Specificity was supported by serologic retesting and virus culture of 11 donors located for follow-up. Sex- and age-specific prevalences did not differ significantly; blacks, however, accounted for 65% of seropositive donations. Within South Florida, one section of Miami had a prevalence of 4.5 per 1,000 donations, significantly above the 0.1 to 1.1 per 1,000 rates for other parts. An epidemiologic association with known HTLV-I endemic areas could account for most infections; all seven typed isolates were characterized as HTLV-I. Exposures, however, were diverse, sometimes multiple, and had no necessary relationship to personal lifestyle. This finding suggests that sources of infection were varied. Seropositive family members emphasize familial clustering of HTLV-I infection.

Prolonged dopamine and serotonin transporter inhibition after exposure to tropanes.

Cocaine and tropane analogs are known to interact with biogenic monoamine transporters by inhibiting amine uptake. Previous in vivo studies have demonstrated that some of these tropanes produce a longer lasting behavioral effect compared with cocaine. We have previously examined several tropane analogs and found a difference in their relative affinities for dopamine (DA) and serotonin (5-HT) transporters. The purpose of this study was to determine the recovery time of transporter function in vitro and in vivo comparing cocaine with the tropane analogs WF-11 (PTT, selective for DA transporters), WF-31 (selective for 5-HT transporters) and WF-23 (highly potent at both DA and 5-HT transporters). In vitro, using primary rat brain cultures of either midbrain or raphe regions, the recovery of the ability to transport either [3H]dopamine or [3H]serotonin, respectively was evaluated at 0, 3, 24, 48, 120 and 240 h after a 1 h exposure to cocaine and tropane analogs. The tropanes exhibited clearance half-lives ranging from 12 to 69 h, while cocaine, on the other hand, exhibited a clearance half-life of approximately 6 h. In studies utilizing [125I]RTI-55 binding, intraperitoneal injections of cocaine and WF-23 into the rat resulted in striatal clearance half-lives ex vivo that were almost identical to those obtained in vitro. These data suggest that the tropanes bind to and reduce transporter function for prolonged periods of time (up to 10-fold longer than cocaine) and those compounds with the highest affinity may produce a pseudo-irreversible inhibition of transporter function.

Release of serotonin induced by 3,4-methylenedioxymethamphetamine (MDMA) and other substituted amphetamines in cultured fetal raphe neurons: further evidence for calcium-independent mechanisms of release.

The substituted amphetamines 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), p-chloro-amphetamine (PCA) and fenfluramine (FEN) all exert their effects by releasing serotonin (5-HT) from presynaptic nerve terminals. In the current study, we examined the ability of these agents to induce the release of 5-HT in culture fetal raphe neurons. The data indicate that the rank order of release potencies for these agents was (+/-)PCA>(+)MDMA=(+)MDA=(+/-)FEN. Studies examining the role fo calcium in 5-HT release demonstrate that preventing calcium influx with L- and N-type calcium channel blockers inhibits potassium-stimulated release of -3H-5-HT but has no effect on release induced by the substituted amphetamines. Furthermore, omitting calcium from the extracellular media or depleting the vesicular pool of neurotransmitter with continual potassium stimulation did not affect the release of -3H-5-HT induced by these compounds. Administration of fluoxetine prior to the substituted amphetamines significantly attenuated the releasing effects of these agents, while producing no effect on potassium-stimulated release. These results are consistent with the notion that the amphetamines induce release of cytoplasmic 5-HT via the plasma membrane transporter.

HIV-1 gp120-induced neurotoxicity to midbrain dopamine cultures.

HIV-1-associated cognitive/motor dysfunction is a frequent neurological complication of acquired immunodeficiency syndrome (AIDS) and has been termed AIDS dementia complex (ADC). The HIV-1 envelope glycoprotein gp120 has been implicated in producing brain injury associated with ADC. The purpose of the present study was to determine if gp120-induced neurotoxicity is associated with damage to dopaminergic systems. Exposure of rat midbrain dopamine cultures to gp120 for 3 days reduced the ability of dopaminergic cells to transport this amine and also resulted in a reduction in dopamine neuron process length while it did not alter either dopamine cell number or the total number of neuronal cells. These detrimental effects of gp120 were prevented by an NMDA receptor antagonist (MK-801) or by preincubation with anti-gp120 antibody. These results suggest that dopaminergic neuronal damage may contribute to the manifestations of AIDS dementia complex.

Methamphetamine-induced alterations in dopamine transporter function.

Repeated methamphetamine (METH) administration has been shown to produce differing neurochemical as well as behavioral effects in rats. This study was designed to examine the effects of acute and chronic METH exposure on uptake and release of [3H]dopamine (DA) in cultured midbrain dopamine neurons to determine if persistent neuronal adaptations ensue. In addition, we have assessed DA D2 receptor function to determine if chronic METH alters this receptor. Fetal midbrain cultures were exposed to METH (1, 10 microM) for 5 days and dopaminergic function examined 1 or 7 days after drug removal. The ability of METH to release [3H]DA was compared to other releasing agents as well as several potent uptake inhibitors. Chronic exposure to a release-promoting concentration of METH resulted in either no change or a reduction in [3H]DA release upon subsequent METH challenge. Pretreatment with METH was also found to cause a decrease in the Bmax for [3H]raclopride binding, suggesting that persistently elevated DA levels cause a downregulation of DA D2 receptors. Examination of transporter kinetics utilizing initial velocity of uptake revealed that METH treatment caused a significant decrease in affinity (K(m)) for the substrate (DA), while not altering the maximal velocity of uptake (Vmax). Binding studies with [125I]RTI-55 revealed that there was no alteration in either the Bmax or Kd for this ligand, suggesting that the changes induced by METH treatment are due to alterations in K(m) and not in the number of DA transport sites. The results from these studies indicate that METH treatment produces a modification in transporter function which may be associated with both the altered uptake and release of [3H]DA. These changes have broad implications for the regulation of transporter activity not only because of the relevance to pre-synaptic mechanisms controlling neurotransmission, but also to the importance of the neuronal adaptation that occurs in response to chronic METH exposure.

First Report of Brown Root Rot on Alfalfa Caused by Phoma sclerotioides in the Continental United States.

Phoma sclerotioides G. Preuss ex Sacc. (previously named Plenodomus meliloti Dearn. & G.B. Sanford) is associated with root rot and extensive winterkill of leguminous forage crops, such as clover (Trifolium and Melilotus spp.), sainfoin (Onobrychis viciifolia), and alfalfa (Medicago sativa). Winterkill and root rot of irrigated alfalfa were observed for the first time in a field of cv. Multiplier in western Wyoming during the spring of 1996. Dark brown to black, sunken, rotting lesions were noted on upper secondary roots and taproots of dead and living diseased plants. Superficial and embedded beaked pycnidia and pycnosclerotia were observed near root lesions. A Phoma sp. isolated from a diseased plant in Farson, WY, was maintained on potato dextrose and half-strength V8-juice agars. Beaked pycnidia, typical of P. sclerotioides, were observed in culture when grown at 10°C for 2 months. A pathogenicity test was performed on cv. Multiplier. Two barley seeds colonized by a Phoma sp. derived from a Wyoming isolate were positioned on taproots of healthy, greenhouse-grown, 5-month-old plants ≈2.5 cm below the crown and were covered with a small piece of sterile cotton. Three replicate samples (24 plants inoculated and 24 plants uninoculated per replicate) were winter-hardened for 4 weeks (15.6°C/10°C, day/night, for 2 weeks, followed by 10°C/7.2°C, day/night, for 2 weeks) and placed outside during January 1998 in Laramie, WY, for a 4-month winter exposure period. Plants were rated for disease during June 1998. A disease severity rating of 1 to 5 was assigned to each experimental unit, where 1 = no disease and 5 = dead plant. The percentage of diseased plants at each severity rating for all inoculated plants was 1 = 19%, 2 = 33%, 3 = 31%, 4 = 13%, and 5 = 4%. Mycelium typical of P. sclerotioides was found on 99% of inoculated plant roots whether or not they had pycnidia. Pycnidia were found on the lower stems and petioles of some inoculated plants. Three percent of control plants also developed brown root rot (BRR) symptoms (taproot lesions or discoloration) by June 1998. The percentage of diseased plants at each severity rating for all uninoculated plants was 1 = 96%, 2 = 4%, and 3 through 5 = 0%. Aboveground propagule placement likely contributed to the spread of BRR by raindrop splash and wind-driven plant debris to adjacent alfalfa. Most inoculated plants had immature pycnidia or protopycnidia (94%), whereas 6.9% of the plants also had fully mature, beaked pycnidia. Pure fungal cultures were obtained from several diseased roots and compared with the original Wyoming Phoma sp. culture and a Canada isolate of P. sclerotioides (ATCC no. 56515) (2): colony, pycnidial, and conidial morphologies were identical, completing Koch's postulates. This is the first report of BRR on alfalfa in the continental United States. References: (1) J. G. N. Davidson. 1990. Brown root rot. Pages 29-31 in: Compendium of Alfalfa Disease. 2nd ed. The American Phytopathological Society, St. Paul, MN. (2) C. R. Hollingsworth et al. Phytopathology 88(suppl.):S39, 1999.

A Rapid Method Using PCR-Based SCAR Markers for the Detection and Identification of Phoma sclerotioides: The Cause of Brown Root Rot Disease of Alfalfa.

A rapid technique for identification and detection of Phoma sclerotioides, the causal agent of brown root rot of alfalfa, has been developed using polymerase chain reaction (PCR). Amplification products obtained from random amplified polymorphic DNA (RAPD) reactions were cloned and sequenced, and two extended primer sets were designed from the resulting data that were used to detect sequence-characterized DNA markers. A single 499-bp DNA amplification product was consistently obtained from primers PSB12499 that was specific for 19 isolates of P.sclerotioides but was not produced from Phoma medicaginis or Phoma betae, or from other soilborne pathogens including Aphanomyces euteiches, Rhizoctonia solani, Fusarium oxysporum, Pythium ultimum, or Phytophthora infestans. A 499-bp amplification product was also produced from root tissue known to be infected with the fungus as verified by microscopic examination. A similar PCR product was obtained from soil samples collected from fields with an established infection of P. sclerotioides on alfalfa. This PCR-based assay enables detection of P. sclerotioides from alfalfa root tissue and in soil samples in a single day, including extraction of DNA, compared with standard methods that require up to 100 days for identification using agar media.

Three-dimensional simulation of lung nodules for paediatric multidetector array CT.

The purpose of this study was to develop and validate a technique for three-dimensional (3D) modelling of small lung nodules on paediatric multidetector array computed tomography (MDCT) images. Clinical images were selected from 21 patients (<18 years old) who underwent MDCT examinations. Sixteen of the patients had one or more real lung nodules with diameters between 2.5 and 6 mm. A mathematical simulation technique was developed to emulate the 3D characteristics of the real nodules. To validate this technique, MDCT images of 34 real nodules and 55 simulated nodules were randomised and rated independently by four experienced paediatric radiologists on a continuous scale of appearance between 0 (definitely not real) and 100 (definitely real). Receiver operating characteristic (ROC) analysis, t-test, and equivalence test were performed to assess the radiologists' ability to distinguish between simulated and real nodules. The two types of nodules were also compared in terms of measured shape and contrast profile irregularities. The areas under the ROC curves were 0.59, 0.60, 0.40, and 0.63 for the four observers. Mean score differences between simulated and real nodules were -8, -11, 13, and -4 for the four observers with p-values of 0.17, 0.06, 0.17, and 0.26, respectively. The simulated and real nodules were perceptually equivalent and had comparable shape and contrast profile irregularities. In conclusion, mathematical simulation is a feasible technique for creating realistic small lung nodules on paediatric MDCT images.

The model repository of the models of infectious disease agent study.

The model repository (MREP) is a relational database management system (RDBMS) developed under the auspices of models of infectious disease agent study (MIDAS). The purpose of the MREP is to organize and catalog the models, results, and suggestions for using the MIDAS and to store them in a way to allow users to run models from an access-controlled disease MREP. The MREP contains source and object code of disease models developed by infectious disease modelers and tested in a production environment. Different versions of models used to describe various aspects of the same disease are housed in the repository. Models are linked to their developers and different versions of the codes are tied to Subversion, a version control tool. An additional element of the MREP will be to house, manage, and control access to a disease model results warehouse, which consists of output generated by the models contained in the MREP. The result tables and files are linked to the version of the model and the input parameters that collectively generated the results. The result tables are warehoused in a relational database that permits them to be easily identified, categorized, and downloaded.

Little Leaguer's shoulder.

A case of Little Leaguer's shoulder in a skeletally immature patient is described with a review of the English literature. This entity manifests as widening of the proximal humeral physis and is well known to our orthopedic colleagues. To our knowledge, however, there is little in the current radiologic literature describing Little Leaguer's shoulder. We describe such a case.

Elimination of "lead time" bias in assessing the effect of early breast cancer diagnosis.

The relationship between diagnosis of breast cancer in the localized stage of disease and improved prognosis for 234 female breast cancer patients aged 55 years and older was examined. Cause of death was used as an indicator of prognosis because, unlike measures of survival, it is unaffected by "lead time" bias. A three- to fourfold increase in the risk of death from breast cancer was observed among women diagnosed in the advanced stages of breast cancer relative to those diagnosed in the localized stage. The results indicate that diagnosis in the localized stage of breast cancer was associated with an improved prognosis, and that "lead time" bias cannot explain the observed association between stage at diagnosis and prognosis for female breast cancer patients aged 55 years and older.