First Report of Rust Caused by Puccinia emaculata on Switchgrass in Arkansas.

In May 2007, switchgrass (Panicum virgatum L.) cv. Alamo and a breeding line, OSU-NSL 2001-1, were planted at the Arkansas Agricultural Research and Extension Center, Fayetteville. In August 2008, a high incidence of dark brown-to-black rectangular foliar lesions delineated by major veins was observed throughout plots of both lines. Lesions covered 25% to nearly 100% of total leaf tissue. Similar symptoms were also observed on unknown switchgrass cultivars in Benton County in northwest Arkansas and in St. Francis County in east-central Arkansas, suggesting that the disease was widely distributed throughout the state. The pathogen produced epiphyllous and adaxial masses of dark brown-to-black telia from erumpent fissures on leaf surfaces. Dark brown teliospores were observed under magnification and were two-celled, oblong to ellipsoid, and 33 ± 3.5 µm long with an apical cell width of 17.5 ± 2.7 µm and basal cell width of 16.2 ± 2.8 µm (reported as mean ± standard deviation, n = 25). Pedicles were colorless to light brown and measured 25.4 ± 9.2 µm (n = 25). In June 2009, at the Fayetteville Research and Extension Center, several second-year stands of switchgrass developed amphigenous and adaxial foliar lesions containing urediniospores. The uredia were globose and finely echinulate, measuring 23.1 ± 2.2 µm (n = 25) with brown cell walls. Teliospore and urediniospore morphology from all collections was consistent with Puccinia emaculata Schw. (2). Genomic DNA was extracted from a representative infected leaf of cv. Alamo, collected in Fayetteville, AR in June 2009, and amplified by PCR with primer sets PRITS1F (3) and ITS4B (1), which amplified an 803-bp fragment of rDNA encoding the first internal transcribed spacer (ITS1), 5.8S subunit, and second internal transcribed spacer (ITS2). The fragment was cloned into pGEM T Easy (Promega Corp, Madison, WI) and sequenced. A BLAST search of GenBank revealed that the fragment was most similar to the rDNA of P. emaculata (GenBank Accession No. EU915294.1; 755 of 758 bases matching; 99% identity) previously reported as a pathogen on switchgrass in Tennessee (3). The incidence and severity of rust on the widely planted switchgrass cv. Alamo is considerable cause for concern as efforts are made to increase acreage and production. Climatic conditions in St. Francis County are generally consistent with locations in Tennessee where switchgrass rust was previously reported (3). However, northwest Arkansas represents the eastern edge of the southwestern United States, suggesting that P. emaculata may affect switchgrass in geographically diverse areas of the United States. To our knowledge, this study represents the first report of rust on switchgrass in Arkansas. Managing this disease will be an important consideration for large-scale switchgrass cultivation in the state. References: (1) M. Gardes and T. D. Bruns. Mol. Ecol. 2:113, 1993. (2) P. Ramachar and G. Cummins. Mycopathol. Mycol. Appl. 25:7, 1965. (3) J. Zale et al. Plant. Dis. 92:1710, 2008.

Impending United States energy crisis.

The U.S. oil and gas industry has been dramatically weakened by the recent oil price collapse. Domestic drilling activity reached a new post-World War II low during the summer of 1986. Given a weak, unstable oil price outlook, U.S. capability will continue to deteriorate. In the last year U.S. imports of foreign oil have risen significantly, and if market forces alone dominate, U.S. dependence is expected to rise from 32% in 1983 to the 50 to 70% level in the not-too-distant future. The 1973 oil embargo and the subsequent attempts to improve U.S. energy security vividly demonstrated the huge costs and long periods of time required to change our energy system. These facts, coupled with the nation's generally short-term orientation, suggest a strong likelihood of a new U.S. energy crisis in the early to middle 1990s.

Catalytic coal gasification: an emerging technology.

Catalytic coal gasification is being developed as a more efficient and less costly approach to producing methane from coal. With a potassium catalyst all the reactions can take place at one temperature, so that endothermic and exothermic reactions can be integrated in a single reactor. A key aspect of the concept involves continuous recycling of product carbon monoxide and hydrogen to the gasifier following separation of methane. Development of the process has advanced steadily since the basic concept was proposed in 1971. A 23-day demonstration run was recently completed in a process development unit with a coal feed rate of 1 ton per day. The next major step in the program will be to design and construct a large pilot plant to bring the technology to commercial readiness in the late 1980's.

The prognostic significance of serum viral load, codon 215 reverse transcriptase mutation and CD4+ T cells on progression of HIV disease in a double-blind study of thymopentin.

OBJECTIVE: To determine in asymptomatic HIV-infected subjects the prognostic value of virion reverse transcriptase (RT) codon 215 mutation, serum HIV RNA level, CD4+ T-cell count and immune complex dissociated (ICD) p24 level. The retrospective evaluation of thymopentin treatment effect on subjects in high risk groups for progression was a secondary objective. PARTICIPANTS: Zidovudine (ZDV)-experienced asymptomatic HIV-infected subjects (n = 352) who had been enrolled in a 48-week placebo-controlled double-blind trial of thymopentin treatment were studied. METHODS: Post hoc analyses were conducted to determine which subjects at study entry were at greater risk for progression to AIDS-related complex (ARC), AIDS or death, and to determine the effect of treatment on these subjects. Four potential prognostic variables (virion RT codon 215 mutation, circulating HIV virion RNA copies, CD4+ T-cell count, and ICD p24) were evaluated by dichotomizing subjects for each variable based on the median of the observed values. CD4+ T-cell count was evaluated prospectively, whereas frozen samples were evaluated under blinded conditions for the other variables after the study was completed. RESULTS: The presence of the codon 215 mutation [P = 0.044; relative hazard (RH), 2.6], > or = 20,000 HIV RNA copies/ml (P = 0.002; RH, 5.5), and < 350 CD4+ cells 10(6)/l (P = 0.042; RH, 2.2) were prognostic factors, and > or = 30 pg/ml ICD p24 level (P = 0.52; RH, 1.4) was not a prognostic factor in predicting progression. Subjects were prestratified by previous ZDV use (< or = 6 or > 6 months). Across both strata thymopentin delayed treatment progression to ARC, AIDS, or death (P = 0.015; RH, 3.0). This effect was magnified in the ZDV-experienced subjects at greater risk, where thymopentin delayed progression compared to placebo in the presence of the codon 215 mutation (P = 0.007; RH, 10.1), > or = 20,000 RNA copies/ml (P = 0.012; RH, 8.9), and CD4+ T-cell count < 350 x 10(6)/l (P = 0.005; RH, 10.4). CONCLUSIONS: Codon 215 mutation, serum HIV RNA and CD4 T-cell count are independent predictors of progression in ZDV-experienced asymptomatic subjects. Furthermore, thymopentin delays HIV disease progression in the presence of a key ZDV resistance mutation as well as high viral load and low CD4+ T-cell counts.

Maintenance of CD4+ cells by thymopentin in asymptomatic HIV-infected subjects: results of a double-blind, placebo-controlled study.

OBJECTIVE: To assess the efficacy and safety of thymopentin in HIV-infected patients who had not yet developed AIDS. DESIGN: Patients were stratified into asymptomatic or symptomatic groups and randomized to receive either thymopentin (50 mg) or placebo, subcutaneously, double-blind for 24 or 52 weeks, three times a week. SETTING: Patients were enrolled at three sites (two hospital clinics and one private practice). PATIENTS: Of 91 HIV-seropositive patients (52 asymptomatic and 39 symptomatic) from whom HIV could be isolated from peripheral blood, 45 were enrolled for 24 weeks and 46 for 52 weeks of double-blind evaluation. MAIN OUTCOME MEASURES: Virological, immunological and clinical evaluations were performed before and during treatment. RESULTS: Thymopentin-treated asymptomatic patients had more CD4+ cells, as demonstrated by a greater area under the percentage CD4+ cells curve (P = 0.03) and a shorter median time to a 20% increase in percentage of CD4+ cells (P = 0.04) in the first 24 weeks, with similar trends in the 52-week study. By 24 weeks no asymptomatic thymopentin-treated and two placebo-treated patients (9.1%, Kaplan-Meier estimate) had progressed to constitutional symptoms (P = 0.12; two-tailed Wilcoxon-Gehan test), with only one further progression in a placebo-treated patient in the subset followed for 52 weeks. Symptomatic patients receiving thymopentin or placebo were similar in both CD4+ cell levels and disease progression (two progressions to AIDS in each group). No serious adverse effects attributable to thymopentin were observed. CONCLUSIONS: These results, if confirmed, indicate that thymopentin, by maintaining CD4+ cells, could slow or arrest immune decline and consequent disease progression at the asymptomatic stage of HIV infection.

Systemic recombinant alpha-2 interferon therapy in relapsing multiple sclerosis.

This report describes the first use of recombinant-DNA-produced human interferon in patients with multiple sclerosis (MS). Ninety-eight patients who were clinically definite for MS with two or more documented exacerbations during the preceding two years were admitted to this placebo-controlled double-blind randomized trial. Although both groups were similar, placebo patients had later MS onset. Patients injected themselves with 2 X 10(6) IU of alpha-2 interferon or placebo three times each week for up to 52 weeks. This dose of interferon was well tolerated in that side effects were minimal. During the trial, the exacerbation rate was sharply reduced in both groups. In the three-month follow-up period after stopping treatment, more patients who were receiving interferon than placebo became worse neurologically. More patients who were receiving interferon than placebo changed from exacerbating MS to progressive MS during the trial. Thus, no clear therapeutic benefit of alpha-2 interferon for MS was detected.

The effect of recombinant alpha 2-interferon on defective natural killer cell activity in multiple sclerosis.

Natural killer (NK) cell activity was evaluated in exacerbating/remitting MS patients. Peripheral blood mononuclear cells (MNC) from MS patients had impaired NK-cell cytotoxicity against the K562 myeloid target cell. NK activity was depressed, irrespective of the interval (2 to 13 months) after the last exacerbation. Recombinant alpha 2-interferon (100 U/ml) enhanced NK activity of both MS and control MNC. Cytotoxicity mediated by interferon-treated MS MNC was increased to the level of untreated control MNC. These studies show that MNC from exacerbating/remitting MS patients possess a defect in NK-cell activity that can be corrected in vitro by treatment with interferon.

Treatment of multiple sclerosis with gamma interferon: exacerbations associated with activation of the immune system.

We treated 18 clinically definite relapsing-remitting MS patients with recombinant gamma interferon in a pilot study designed to evaluate toxicity and dosage. Patients received low (1 microgram), intermediate (30 micrograms), or high (1,000 micrograms) doses of interferon by intravenous infusion twice a week for 4 weeks. Serum levels of gamma interferon were proportional to dose and no interferon was detected in CSF. Seven of the 18 patients had exacerbations during treatment, a significant increase compared with the prestudy exacerbation rate (p less than 0.01). Exacerbations occurred in all three dosage groups and were not precipitated by fever or other dose-dependent side effects. There were significant increases in circulating monocytes bearing class II (HLA-DR) surface antigen, in the proliferative responses of peripheral blood leukocytes, and in natural killer cell activity. These results show that systemic administration of gamma interferon has pronounced effects on cellular immunity in MS and on disease activity within the CNS, suggesting that the attacks induced during treatment were immunologically mediated. Gamma interferon is unsuitable for use as a therapeutic agent in MS. Agents that specifically inhibit gamma interferon production or counteract its effects on immune cells should be investigated as candidates for experimental therapy.

Vaccine-associated poliomyelitis in a household contact with Netherton's syndrome receiving long-term steroid therapy.

Paralytic poliomyelitis developed in a man 51 days after his two-month-old daughter received her first dose of trivalent live oral poliovirus vaccine. The patient was receiving long-term glucocorticosteroid therapy (tapered to 12.5 mg per day for the eight months prior to his poliomyelitis) for Netherton's syndrome, a congenital syndrome characterized by bamboo-like hair, hyperkeratotic and hyperhidrotic skin, and multiple allergies. The patient was ventilator-dependent and quadriplegic throughout most of his hospital stay and died in the hospital 10 months after the onset of paralysis. "Vaccine-like" type 3 poliovirus was isolated from a stool specimen and his serum showed a significant rise in neutralizing antibody titer against type 3 virus. This case report represents the first documented case of vaccine-associated poliomyelitis in a household contact receiving glucocorticosteroids, although evidence of immunosuppression was not documented. Nevertheless, the case reinforces current recommendations not to administer oral poliovirus vaccine to persons known to be immune deficient or suppressed or to normal persons with close contacts known to be immune deficient or suppressed.

T8 antigen density on peripheral blood lymphocytes remains unchanged during exacerbations of multiple sclerosis.

Fluorescence-activated cell sorter (FACS) analysis was used to quantitate the antigen density of the lymphocyte markers T3, T4, T8, and Leu 7 on the surface of peripheral blood mononuclear cells (MNC) from exacerbating/remitting multiple sclerosis (MS) patients. Serial studies showed that the mean fluoresce intensity for all the markers studied did not change significantly during exacerbations. There was a tendency for a generalized decline in T-cells, evidenced by a drop in both T8+- and T4+-cells; however, these changes were not statistically significant. It appears, therefore, that the lymphocyte markers studied here are not useful as markers of disease activity in exacerbating/remitting MS.

Development of age-dependent resistance to sindbis virus encephalitis: correlation with inactivation of virus within the blood stream.

The clearance of Sindbis virus from the blood stream and its localization in the reticuloendothelial system (RES) was studied in mice susceptible (2 weeks old) and resistant (6 weeks old) to fatal Sindbis virus encephalitis. No significant differences in the relative capacity of 2-week-old and 6-week-old mice to remove 125I-labelled virus from the blood stream and to localize virus within the liver were observed. However, the decline of infectious virus was more rapid in the blood of adult mice. These studies suggest that, in addition to physical removal of virus by the RES, inactivation of virus in the blood serum stream plays an important role in limiting viremia during infections with Sindbis virus.

Respiratory syncytial virus immune globulin treatment of lower respiratory tract infection in pediatric patients undergoing bone marrow transplantation - a compassionate use experience.

Respiratory syncytial virus (RSV) pneumonia in BMT recipients carries a mortality rate of approximately 50-70% despite ribavirin (Virazole) treatment. In both immunocompetent and immunocompromised animal models, RSV neutralizing antibodies rapidly reduce pulmonary virus load after a single dose. RSV-IGIV (RespiGam) is an IgG immune globulin with high concentrations of RSV neutralizing antibody (>19 200 MU/ml). From June 1991 to February 1996, a compassionate-use protocol using RSV-IGIV for treatment of RSV infections was conducted. Eleven children at multiple centers, mean age 3.3 years (4 months to 9 years), were undergoing BMT and met the protocol criteria. They received a single 1500 mg/kg dose of RSV-IGIV infused over 12 h at a median of 5 days (1-37 days) after RSV symptom onset. Ten of these patients received prior or concurrent aerosolized ribavirin. Serum RSV neutralizing titers were measured in five patients and showed a 3- to 30-fold increase 24 h after RSV-IGIV infusion. Adverse events were mild. One of 11 (9.1%) patients died from their RSV illness (91% RSV survival). In comparison to previously published reports, RSV-IGIV treatment of RSV pneumonia in BMT patients may increase survival above that in such patients treated with ribavirin alone. Bone Marrow Transplantation (2000) 25, 161-165.

Immunosuppression in goats inoculated with parainfluenza type 3 virus.

The humoral and cellular immune responses of goats experimentally infected with an ovine isolate of parainfluenza type 3 virus (PI-3) were examined. Virus neutralization and enzyme-linked immunosorbent assays were used to determine antibody in the serum and CSF. Lymphocyte stimulation, measured by [3H]thymidine incorporation into peripheral blood leukocytes, was used to determine cellular responses to phytomitogens and virus. There were significant suppression of peripheral blood leukocyte responses to T-cell mitogens early in the course of infection and delayed onset of virus-specific cell-mediated immunity. Delay in antibody formation did not occur. The suppression of mitogen response has been reported with other paramyxovirus infections. The importance of the suppressed cellular immune response for potentiating other infective agents is discussed.

Entry overload, emergency department overcrowding, and ambulance bypass.

OBJECTIVES: To describe an experience of emergency department (ED) overcrowding and ambulance bypass. METHODS: A prospective observational study at Royal Perth Hospital, a major teaching hospital. Episodes of ambulance bypass and their characteristics were recorded. RESULTS: From 1 July 1999 to 30 June 2001, there were 141 episodes of ambulance bypass (mean duration 187 min, range 35-995). Monday was the most common day with 39 (28%) episodes. Entry block alone was the most common reason bypass was activated (n=38, 30.4%). The mean number of patients in ED at these times was 40 (occupancy 174%), including nine in the corridor, seven awaiting admission, and 14 waiting to be seen. Episodes attributable to entry block were typically preceded by a presentation rate of >/=10 patients per hour for >/=2 hours (OR 6.2, 95% CI 4.3 to 8.5). Mid-afternoon to early evening was the most common time for activation. Ambulance bypass is increasing in frequency and duration. CONCLUSIONS: Entry overload resulting in entry block results from overwhelming numbers of patients presenting to the ED in a short space of time. Entry block impairs access to emergency care. Unless something is done in the near future, the general public may no longer be able to rely on EDs for quality and timely emergency care. A "whole of system" approach is necessary to tackle the problem.

Identifying genes in Fusarium verticillioides through forward and reverse genetics.

The increasing availability of sequenced genomes for plant pathogenic fungi has revolutionized molecular plant pathology in recent years. However, the genetic regulatory networks underlying many important components of pathogenesis remain poorly defined. Although the protocols outlined in this chapter can be utilized to identify genes regulating a wide range of biological processes in many filamentous fungi, we focus on describing how to identify genes through forward and reverse genetics, using the plant pathogenic fungus Fusarium verticillioides as a model for the protocol. Specifically, this chapter explains how to create a collection of insertional mutants via Restriction Enzyme Mediated Integration (REMI) and how to screen mutants with a high-throughput method to visualize defects in amylolysis. Next, techniques are described to define the genomic lesions in REMI mutants with genome-walker PCR in order to identify candidate genes. Finally, protocols are presented describing a reverse-genetic approach to disrupt candidate genes in the wild-type strain with a split-marker strategy to confirm the phenotype observed in the REMI mutant.