First Report of Box Blight Caused by Cylindrocladium pseudonaviculatum (C. buxicola) in British Columbia, Canada.

Boxwoods (Buxus spp.) are common woody ornamental hedging plants in Europe and North America, typically propagated by cuttings. In October 2011, shoot dieback and defoliation was observed on Buxus sempervirens 'Suffruticosa' (dwarf English boxwood) and 'Green Balloon' in outdoor, 10-cm pots at a wholesale nursery in Chilliwack, British Columbia. Circular leaf spots with black rings occurred on leaves and black, water-soaked, cankers girdled the stems and petioles. Leaf and stem samples were collected on November 21, 2011, and incubated for 48 h in a moist chamber at room temperature. In addition to Volutella buxi, a Cylindrocladium species producing conidia on white sporodochia was observed on host tissue under the microscope. Leaves with lesions were surface-sterilized in 10% bleach for 30 to 60 s, rinsed in sterile water, and lesions were cut out and plated on PDA and carnation leaf media. The species was identified as Cylindrocladium pseudonaviculatum Crous, J.Z. Groenew. & C.F. Hill 2002 by comparison of conidia and phialide morphology to published descriptions. Conidia were hyaline, one-septate, cylindrical with rounded ends and 38 to 76 µm (mean 51 µm) × 4 to 6 µm on carnation leaf media and 41 to 66 µm (mean 52 µm) × 4 to 6 µm on B. sempervirens 'Suffruticosa' leaves, comparable to the reported range of 40 to 75 × 4 to 6 µm (1,2,3,4). Conidia were produced in clusters on terminal, ellipsoid vesicles at the tips of penicillate conidiophores. Vesicles were 10.2 (7.6 to 12.8 µm) at the widest point, consistent with the 6 to 11 µm reported in (2,3) and tapered to a rounded point; stipe extensions were septate and measured an average of 130 µm (107 to 163 µm) in length to the tip of the vesicle, consistent with the 95 to 155 µm reported in (1), 89 to 170 µm reported in (2), and 95 to 165 µm in (3). Chlamydospores were not observed on host tissue but appeared in older PDA cultures as dark brown microsclerotia. DNA was extracted from single-spore colonies on PDA and the internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1 and ITS4. The ITS sequence (GenBank Accession No. KC291613) was 100% identical to C. buxicola strain CB-KR001 (HM749646.1) and Calonectria pseudonaviculata strain ATCC MYA-4891 (JX174050.1). In early December 2011, box blight was identified on container-grown B. sinica var. insularis × B. sempervirens 'Green Velvet,' 'Green Gem', and 'Green Mountain' and B. sempervirens L. (common or American boxwood). The pathogen was identified by microscopic examination at three wholesale nurseries in the eastern Fraser Valley and one landscape planting. The isolate has been deposited in the Canadian Collection of Fungal Cultures in Ottawa, Canada (DAOM 242242). References: (1) B. Henricot and A. Culham. Mycologia 94:980, 2002. (2) K. L. Ivors, et al. Plant Dis. 96:1070, 2012. (3) C. Pintos Varela, et al. Plant Dis. 93:670, 2009. (4) M. Saracchi, et al. J. Plant Pathol. 90:581, 2008.

First Report of Blueberry scorch virus in Black Huckleberry in British Columbia.

Blueberry scorch virus (BlScV), an aphid-borne carlavirus, causes a serious disease of highbush blueberry (Vaccinium corymbosum L.) in North America and Europe. Symptoms of BlScV infection on highbush blueberry include necrosis of flower blossoms and young leaves, shoot blight, and chlorosis. Currently, cranberry (Vaccinium macrocarpon L.) is the only other natural host of BlScV. In July 2004, wild black huckleberry (Vaccinium membranaceumL.) was sampled in the Kootenay Region of southeastern British Columbia. Foliar tissues were sampled during 2004 from 11 bushes from a clearing on the side of a mountain near Crawford Bay, BC, Canada and tested by double-antibody sandwich-ELISA using polyclonal antiserum (Agdia Inc., Elkhart, IN). BlScV was detected in 6 of the 11 bushes sampled and in the positive control (BlScV-infected blueberry leaf tissue) and was not detected in the negative control (healthy blueberry leaf tissue). To confirm the presence of the virus, total nucleic acid was extracted from ELISA-positive huckleberry samples according to an established protocol (A. Rowhani et al. Proc. Int. Counc. Stud. Viruses Virus-Like Dis. Grapevine, Extended Abstr. 13:148, 2000). Reverse transcription-PCR was performed using pd(T)12-18 random primer (Amersham Biosciences, Piscataway, NJ) for reverse transcription and BlScV-specific primers developed against the published NJ-2 sequence of BlScV (GenBank Accession No. NC_003499). Using the forward primer, BS708F, (5'-TCAATCCGTGGTGCTACGAG-3'), and the reverse primer, BS1188R, (5'-ACAGTGCGCAATGTTCCAGT-3'), a 480-bp amplicon was obtained from each of the ELISA-positive samples, while no ampli-cons were observed for the negative control (ELISA-negative huckleberry tissue). Direct sequencing of one selected amplicon revealed 90, 84, and 77% nucleotide sequence identity and 97, 96, and 88% amino acid sequence identity with strains NJ-2, BC-1 (GenBank Accession No. AY941198) and BC-2 (GenBank Accession Nos. AY941199), respectively. BlScV-infected huckleberries were asymptomatic. The presence of BlScV in alternate hosts has implications for disease epidemiology. Testing for BlScV in Vaccinium species in and around commercial highbush blueberry plantings, as well as lowbush blueberry (V. angustifolium Aiton), rabbiteye blueberry (V. ashei Reade), other native Pacific Northwest species (V. ovatum Pursh and V. parvifolium Smith), and ornamental Vaccinium species is warranted. To our knowledge, this is the first report of BlScV infecting black huckleberry.

First Report of Blueberry scorch virus in Cranberry in Canada and the United States.

Blueberry scorch disease, caused by the carlavirus Blueberry scorch virus(BlScV), is a serious disease of highbush blueberry (Vaccinium corymbosum L.) in North America and Europe. Symptoms of BlScV infection in highbush blueberry include necrosis of flower blossoms and young leaves, shoot blight, and chlorosis. In June 2003, BlScV was detected for the first time in cranberry (Vaccinium macrocarpon L.) in Abbotsford, British Columbia, Canada using double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Ten uprights were sampled from different locations in the bog, seven of which tested positive using DAS-ELISA. To confirm infection, total nucleic acid was extracted from infected cranberry leaf tissue according to an established protocol (2). Primers were developed against the published NJ-2 sequence of BlScV (GenBank Accession No. NC_003499). The forward primer, BS4586F (5'-CTTCAGGATGAGTGGTGCAA-3'), and the reverse primer, BS5164R (5'-CGCGTGCTGGAAGCATACAA-3'), were used in reverse-transcription polymerase chain reaction (RT-PCR) to amplify a portion of the RNA-dependent RNA polymerase gene of BlScV. The amplicons of the expected size were sequenced and the nucleotide sequence of the product showed 82% homology, and the predicted amino acid sequence had 91% homology with the published NJ-2 sequence. A random survey for BlScV in cranberry bogs in the Pacific Northwest Region of North America was conducted. Testing revealed the presence of BlScV in 7 of 42 bogs in British Columbia, Canada, 2 of 12 bogs in Oregon, and 3 of 18 bogs in Washington. Nucleotide sequencing of RT-PCR products of BlScV from a Washington bog showed 87 and 96% homology at the nucleotide and predicted amino acid level, respectively, in the methyltransferase of the NJ-2 strain using forward primer, BS276F (5'-CCGTCTGCAAGACATTAGAG-3'), and reverse primer, BS743R (5'-TCTTCTTCACCTCGTACTCG-3'). Several bogs had a high incidence of BlScV infection, with over 70% of the sampled uprights testing positive. Some infected cranberry bogs were adjacent to infected blueberry fields, while others were isolated. Transmission of the virus is likely to have occurred by aphids, which are known to be vectors of BlScV (1). Presently, BlScV appears to be asymptomatic in cranberry. To our knowledge, this is the first report of BlScV infecting cranberry. The potential for infection of other Vaccinium spp., such as lowbush blueberry (V. angustifolium Aiton), rabbiteye blueberry (V. ashei Reade), native Pacific Northwest species (V. membranaceum Douglas ex Torrey, V. ovatum Pursh, and V. parvifolium Smith), and ornamental Vaccinium spp. needs to be investigated. References: (1) P. R. Bristow et al. Phytopathology 90:474, 2000. (2) D. W. Hughes and G. Galau. Plant Mol. Biol. Rep. 6:253, 1988.

Effect of chromatic dispersion on nonlinear phase noise.

We consider the combined effects of amplified spontaneous emission noise, optical Kerr nonlinearity, and chromatic dispersion on phase noise in an optical communication system. The effect of amplified spontaneous emission noise and Kerr nonlinearity were considered previously by Gordon and Mollenauer [Opt. Lett. 15, 1351 (1990)], and the effect of nonlinearity was found to be severe. We investigate the effect of chromatic dispersion on phase noise and show that it can either enhance or suppress the nonlinear noise amplification. For large absolute values of dispersion the nonlinear effect is suppressed, and the phase noise is reduced to its linear value. For a range of negative values of dispersion, however, nonlinear phase noise is enhanced and exhibits a maximum related to the modulation instability found in amplitude fluctuations. Nonlinear phase noise is quenched by these effects even in dispersion-compensated systems; the degree of suppression is sensitively dependent on the dispersion map. We demonstrate these results analytically with a simple linearized model.

Schrödinger equation with a spatially and temporally random potential: effects of cross-phase modulation in optical communication.

We model the effects of cross-phase modulation in frequency (or wavelength) division multiplexed optical communications systems, using a Schrödinger equation with a spatially and temporally random potential. Green's functions for the propagation of light in this system are calculated using Feynman path-integral and diagrammatic techniques. This propagation leads to a non-Gaussian joint distribution of the input and output optical fields. We use these results to determine the amplitude and timing jitter of a signal pulse and to estimate the system capacity in analog communication.

Comparison of agar dilution, broth microdilution, E-test, disk diffusion, and automated Vitek methods for testing susceptibilities of Enterococcus spp. to vancomycin.

An evaluation was undertaken to determine the optimal method for testing the susceptibilities of 100 clinical isolates and two reference strains of Enterococcus spp. to vancomycin in vitro. Six testing methods were studied by using the following media and incubation times: agar screen with the Synergy Quad Plate (Remel, Lenexa, Kans.), an in-house-prepared brain heart infusion (BHI) agar plate, and an in-house-prepared Mueller-Hinton (MH) agar plate, all incubated for 24 or 48 h; broth microdilution (Sensititre Just One Strip; AccuMed International, Inc., West Lake, Ohio) with BHI or cation-adjusted MH broth incubated for 24 or 48 h; agar dilution with BHI or MH agar incubated for 24 or 48 h; epsilometer test (E test; AB BioDisk, Solna, Sweden) with BHI or MH agar incubated for 24 or 48 h; disk diffusion with BHI or MH agar incubated for 24 or 48 h; and the automated Vitek method with the gram-positive susceptibility Staphylococcus aureus card and R02.03 software (bioMerieux, Inc., Hazelwood, Mo.). Growth failures occurred with MH media (n = 6) but not with BHI media. One growth failure occurred with the Vitek method. Results for each testing method for each Enterococcus strain were interpreted as susceptible, intermediate, or resistant according to current National Committee for Clinical Laboratory Standards (NCCLS) criteria and compared to the vancomycin resistance genotype (i.e., vanA, vanB, vanC-1, or vanC-2/3). For all methods, extension of the incubation time from 24 h to 48 h either produced no difference in the results or gave poorer results. The following methods produced no very major or major interpretive errors: broth microdilution with BHI media incubated for 24 h, agar dilution with BHI media incubated for 24 or 48 h, and E test with BHI media incubated for 24 or 48 h. Unacceptable frequencies of very major errors (> 1%) occurred with all methods for which MH media were used. Minor interpretive errors were frequent with all methods. These minor interpretive errors also occurred most frequently with Enterococcus strains with vanC genes, which encoded low-level vancomycin resistance (MIC < or = 8 microg/ml), as opposed to Enterococcus strains which possessed vanA or vanB genes, which encoded higher-level vancomycin resistance (MIC > or = 64 microg/ml). Modification of NCCLS breakpoints, especially for motile Enterococcus spp. (E. casseliflavus, E. flavescens, and E. gallinarum), may resolve this problem; however, in the current study, one E. faecalis strain and one E. faecium strain carried only the vanC gene. The agar screen method may also require reformulation. The current agar screen plate contains 6 microg of vancomycin per ml, which may not detect all low-level resistance associated with vanC genotypes. Nevertheless, the clinical significance of this low-level vancomycin resistance remains unknown.

Static and dynamic balance responses in persons with bilateral knee osteoarthritis.

Research has documented decreased strength and proprioception in people with arthritis. Both are components of balance, but reliable studies documenting balance deficits have not been done. This study tested standing balance in an adult population with osteoarthritis. The osteoarthritic group (N = 11) and the age-matched control group (N = 10) were tested on the Balance System. Each underwent two trials of six testing conditions [two visual conditions (eyes open, eyes closed) under each of three platform conditions (stable, angular rotation, and linear translation)] and completed a functional assessment scale. Individuals with knee osteoarthritis demonstrated significantly more postural sway than the control group across conditions (p < 0.02). The functional assessment scale developed to discriminate between the two groups demonstrated internal consistency (Cronbach's coefficient alpha = 0.83), and scores were significantly different between the two groups (p < 0.0001). Results suggest the importance of balance training in this population. The functional assessment scale may be a useful tool to document functional levels in knee osteoarthritis.

Comparison of the pathology of fascia in eosinophilic myalgia syndrome patients and idiopathic eosinophilic fasciitis.

The L-tryptophan eosinophilic myalgia syndrome (EMS) clinically has some similarities with idiopathic eosinophilic fasciitis (EF). In order to study the pathology of both syndromes, we analyzed 21 biopsies of patients with EMS and 8 with idiopathic EF. In both diseases there is dermal and fascial mucin and dermal edema, but this was more common in EMS. EMS is also characterized by dilated lymphatics, dermal and septal sclerosis and macrophage-rich inflammation. Neural inflammation was seen in 4 of the cases with EMS and in none with idiopathic EF. In both syndromes, there are many histopathological similarities. The differences may be due to sampling and to sample size. The nerve lesions of EMS may result from the nature of lymphocyte-macrophage inflammation, or the effect of the eosinophil neurotoxin and may not be a primary event.

Acute intravenous calcitonin: failure to modify systemic blood pressure.

Calcium and a principal calcium-regulating hormone, PTH, have been characterized as possessing vasoactive properties in the spontaneously hypertensive rat (SHR). Calcitonin is another calcium-regulating peptide with primary, but opposing effects on many of the same target organs, and capable of modifying both extracellular and intracellular calcium distribution. We sought to determine whether calcitonin, like PTH, exhibits vasoactivity in the SHR and its control, the Wistar-Kyoto rat (WKY). Three male SHR and 3 male normotensive WKY received intravenous injections (range 1-100 micrograms/kg) of synthetic human calcitonin. Seven SHR and 7 WKY received equivalent doses of the more potent peptide, synthetic salmon calcitonin. Intraarterial pressure was monitored continuously. Neither analog of calcitonin produced significant changes in blood pressure. Serum ionized calcium levels 30 minutes postinjection were unchanged from baseline in the WKY; in the SHR, postinjection serum ionized calcium levels were significantly lower than baseline values (pre = 1.12 +/- 0.01 mmol/l vs. post 1.08 +/- 0.01 mmol/l, p less than 0.05). We conclude that calcitonin modifies extracellular calcium, but does not have demonstrable, acute systemic cardiovascular effects.

Intestinal basement membrane of Ascaris suum. Characterization of carbohydrate units.

Characterization of glycopeptides obtained on alkaline hydrolysis and on extensive collagenase and pronase digestion of the intestinal basement membrane showed the existence of two distinctly different carbohydrate units. One of these is a disaccharide, composed of glucose and galactose, linked to hydroxylysine. It was shown to be identical to the unit (2-O-D-glucopyranosyl-O-D-galactopyranosylhydroxylyasine) present in vertebrate basement membranes, as determined from stability to alkaline hydrolysis, retention time on amino acid analyzer, chemical composition, graded acid hydrolysis, methylation analysis, and periodate oxidation. Direct quantitation after alkaline hydrolysis showed the presence of 9.71 disaccharide units/1000 amino acid residues, indicating that 89% of the hydroxylysine residues of the intestinal membrane are glycosylated. The other unit, consisting of the remaining monosaccharides of the membrane, was separated from the disaccharide unit by gel filtration and ion exchange chromatography of collagenase/pronase digests. Chemical analyses and molecular weight determination by thin layer gel filtration chromatography of purified glycopeptides indicated that this unit is an oligosaccharide which is composed of fucose, galactose, mannose, galactosamine, and glucosamine in a mole ratio of 1:1:1:1:2, respectively. The amount of this unit was calculated to be 2.6 units/1000 amino acid residues.

Procoagulant activity of platelets.

A simplified, semiautomated, quantitative method to evaluate platelet procoagulant activity was evaluated on plasma that contained widely divergent concentrations of platelets. When fewer than 100,000 platelets/cu mm were present, excellent correlation between platelet concentration and clotting time was noted, with correlation best in the range 30,000 to 100,000 platelets/cu mm. When platelets were present in concentrations greater than 100,000/cu mm, no alteration in clotting time as a function of platelet count was noted. These observations were consistent with clinical observations that patients with greater than 100,000 platelets rarely bleed and that those with fewer than 30,000 platelets bleed in an unpredictable fashion. The procedure appears to be of potential value in evaluation of patients for defects in platelet factor 3 release.

Spontaneous platelet aggregation. Occurrence in an asymptomatic individual.

Spontaneous platelet aggregation in an asymptomatic individual is described. The platelet-poor plasma of the subject greatly enhanced the response of normal platelet-rich plasma to adenosine diphosphate. The spontaneous platelet aggregation was easily inhibited by aspirin.