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Dengue virus infects approximately 100 million people annually, but there is no available therapeutic treatment. The mimetic peptide, DN59, consists of residues corresponding to the membrane interacting, amphipathic stem region of the dengue virus envelope (E) glycoprotein. This peptide is inhibitory to all four serotypes of dengue virus, as well as other flaviviruses. Cryo-electron microscopy image reconstruction of dengue virus particles incubated with DN59 showed that the virus particles were largely empty, concurrent with the formation of holes at the five-fold vertices. The release of RNA from the viral particle following incubation with DN59 was confirmed by increased sensitivity of the RNA genome to exogenous RNase and separation of the genome from the E protein in a tartrate density gradient. DN59 interacted strongly with synthetic lipid vesicles and caused membrane disruptions, but was found to be non-toxic to mammalian and insect cells. Thus DN59 inhibits flavivirus infectivity by interacting directly with virus particles resulting in release of the genomic RNA.

Abstract Viral fusion proteins mediate cell entry by undergoing a series of conformational changes that result in virion-target cell membrane fusion. Class I viral fusion proteins, such as those encoded by influenza virus and human immunodeficiency virus (HIV), contain two prominent alpha helices. Peptides that mimic portions of these alpha helices inhibit structural rearrangements of the fusion proteins and prevent viral infection. The envelope glycoprotein (E) of flaviviruses, such as West Nile virus (WNV) and dengue virus (DENV), are class II viral fusion proteins comprised predominantly of beta sheets. We used a physio-chemical algorithm, the Wimley-White interfacial hydrophobicity scale (WWIHS) 1 in combination with known structural data to identify potential peptide inhibitors of WNV and DENV infectivity that target the viral E protein. Viral inhibition assays confirm that several of these peptides specifically interfere with target virus entry with 50% inhibitory concentration (IC50) in the 10 μM range. Inhibitory peptides similar in sequence to domains with a significant WWIHS scores, including domain II (IIb), and the stem domain, were detected. DN59, a peptide corresponding to the stem domain of DENV, inhibited infection by DENV (>99% inhibition of plaque formation at a concentrations of 99% inhibition at

Phytophthora root rot on Abies fraseri trees was monitored from 2001 to 2007 within the disease front of a 12-year-old Virginia plantation where trees had been dying of the disease since 1994. After a slow increase in early foliage symptom development from July 2001 to September 2002, the frequency of A. fraseri trees with early symptoms accelerated for about 15 months. While the slow increase occurred during a 18.7% lower than normal rainfall period and the acceleration occurred during a 31.2% higher than normal rainfall period, the percentage of trees with early symptoms continued to increase during the mid-winter months (December–February) when the estimated mean minimum daily soil temperature (25 cm depth) was unfavourable (<10°C) to Phytophthora cinnamomi pathogenic activity. The time required for trees to progress from early foliage symptoms to completely dead foliage, from November 2000 to October 2007, was highly variable, ranging from 4 to 35 months. Root recovery rates for P. cinnamomi, assayed on a selective medium, were 6.4 times greater for symptomatic foliage trees than for asymptomatic foliage trees in this deep, silt-loam soil. Following an atypical cold period in February 2007, when the mean minimum daily soil temperature was 0.8°C, symptomatic roots yielded only a low level of germinable propagules of P. cinnamomi. Further, during an atypical midsummer in 2007 (June–August), when the soil water potential was at or below −9 bars for 68 of 92 days, symptomatic roots yielded no germinable propagules of P. cinnamomi. Addition of thiophanate-methyl to the selective medium aided P. cinnamomi isolation by inhibiting many undesired pythiaceous colonies growing from symptomatic roots. [ABSTRACT FROM AUTHOR]

Objectives: The purpose of this investigation was to determine the feasibility of using the polymerase chain reaction to detect bacteria in amniotic fluid and to compare pregnancy outcomes in subsets of women categorized by amniotic fluid culture, polymerase chain reaction, and interleukin-6 findings.Study Design: Amniotic fluid from 54 pregnancies with preterm labor and no clinical evidence of intraamniotic infection was evaluated with use of the polymerase chain reaction, interleukin-6, and bacterial culture. Gestational age, newborn weight, and time between amniocentesis and delivery were compared between subsets of women categorized by these tests.Results: With use of the polymerase chain reaction <100 bacteria per milliliter could be detected in amniotic fluid. A total of 55.5% of the amniotic fluid samples were polymerase chain reaction positive, whereas 9.2% of culture results were positive. Birth weights and gestational age at delivery were less and time from amniocentesis to delivery was shorter in the polymerase chain reaction-positive group (p < 0.05). Nine samples (15%) had elevated interleukin-6 concentrations; of these, six were polymerase chain reaction positive.Conclusions: The polymerase chain reaction is a sensitive means of detecting bacteria in amniotic fluid. These results provide further evidence of an association between preterm delivery and intraamniotic infection. Not all amniotic fluid samples with elevated interleukin-6 levels have bacteria detectable by the polymerase chain reaction. We anticipate that the polymerase chain reaction will provide another avenue for the detection of bacteria in amniotic fluid. [ABSTRACT FROM AUTHOR]

Twenty-four healthy Holstein calves received single subcutaneous injections of 2.5 mg/kg bw tulathromycin. At each of 8 time points ranging from 1 - 360 hr post-dose, three animals were randomly selected for euthanasia. At the time of euthanasia, whole blood, lung tissue, and broncho-alveolar lavage (BAL) samples were collected and subsequently assayed for tulathromycin with an LC MS/MS procedure. Concentrations of tulathromycin in pulmonary epithelial lining fluid (PELF) were determined from the BAL supernatant concentrations using the urea dilution method. Concentrations of tulathromycin in the cells of the pulmonary epithelial lining fluid (PELF cells, predominately macrophages) were also determined. Tulathromycin distributed rapidly and extensively into PELF, PELF cells, and lung homogenate, with areas under the concentration- time curve through 360 hr (AUC0-360) that were 53 - 565 times the plasma AUC0-360. Tulathromycin concentrations exceeded 1,000 ng/mL in PELF and 1,000 ng/g in lung homogenate by 3 hr post-dose, and the peak concentration in BAL cells was 19,500 ng/mL at 72 hr. The drug was slowly eliminated from lung homogenate, PELF, and PELF cells, with apparent t1/2 values >200 hr. These data demonstrate rapid achievement and prolonged maintenance of therapeutic tulathromycin concentrations at sites of respiratory infections in cattle following a single-dose administration. [ABSTRACT FROM AUTHOR]