First Report of Verticillium Wilt of Ailanthus altissima in Virginia Caused by Verticillium nonalfalfae.

Ailanthus altissima (Mill.) Swingle, commonly known as tree-of-heaven, is an invasive tree species that has spread throughout the United States since its introduction in 1784 (2). During a survey in July 2009, approximately 1,100 A. altissima trees were observed at two locations in western Virginia (a roadside in Montgomery Co. and a wooded area adjacent to a railroad in Bedford Co.) exhibiting foliar wilt symptoms, defoliation, yellowish vascular discoloration, or death at an incidence of ~77%. Similar symptoms on A. altissima were reported in Roanoke, VA in the early 1930s and after 2005 in Pennsylvania, attributed to a Verticillium sp. (1,2). To identify the causal agent, discolored xylem tissue samples were excised from 10 symptomatic A. altissima trees at both locations, soaked in 1% NaOCl for 2 min, rinsed with sterilized distilled water for 5 min, and placed onto plum extract agar. Cultures were incubated in the dark at 22°C for 7 to 14 days. The resultant colonies (three to four per location) were subcultured and identified putatively as a Verticillium sp. closely related to Verticillium albo-atrum Reinke and Berthold (3), based on melanized, thick-walled, resting mycelia and phialides arranged in verticillate whorls that amassed round, oval-shaped conidia (5.1 ± 1.2 µm × 2.8 ± 0.4 µm, n = 100). Molecular identification of two fungal isolates (one per location) was determined by amplification of the protein coding genes elongation factor 1-alpha (EF), glyceraldehyde-3-phosphate dehydrogenase (GPD), and tryptophan synthase (TS), using PCR primers developed recently for Verticillium (3). A BLAST search on the edited contigs revealed 100% sequence similarity for all three protein coding genes among the two isolates and reference sequences of isolates PD592 (GenBank Accessions JN188227, JN188163, and JN188035 for EF, GPD, and TS, respectively) and VnAaPA140 (KC307764, KC307766, and KC307768 for EF, GPD, and TS, respectively) of the newly-proposed species, V. nonalfalfae (formerly V. albo-atrum). Aligned sequences from one representative isolate, VnAaVA2 (Bedford Co.), were deposited into GenBank as KC307758 (EF), KC307759 (GPD), and KC307760 (TS). To confirm pathogenicity to A. altissima, the two molecularly characterized isolates (one per location) were inoculated into 18 10-week old A. altissima stems that were grown in an environmental chamber at 24°C, 60% RH, and a 12-h photoperiod from seeds collected in Blacksburg, VA. A conidial suspension of each isolate was injected into each stem (0.1 ml of 1 × 108 CFU/ml/stem). All 36 seedlings inoculated with the proposed V. nonalfalfae isolates developed wilting of leaflets within 2 weeks post-inoculation (WPI), defoliation of leaflets by 6 WPI, and were dead by 9 WPI. Eighteen control seedlings were inoculated similarly with distilled water, and remained asymptomatic. Fungi resembling the proposed species V. nonalfalfae were reisolated from all inoculated stems except the control plants, and the species confirmed morphologically as described above. V. nonalfalfae is a recently proposed species that can infect a variety of plant species (3). To our knowledge, this is the first report of this proposed species on A. altissima in Virginia. New state reports of this pathogen on A. altissima are important for regulatory issues associated with using this pathogen as a potential biological control agent. References: (1) G. F. Gravatt and R. B. Clapper. Plant Dis. Rep. 16:96, 1932. (2) M. J. Schall and D. D. Davis. Plant Dis. 93:747, 2009. (3) P. Inderbitzin et al. PLoS ONE, 6, e28341, 2011.

Host-range testing of Eucryptorrhynchus brandti (Coleoptera: Curculionidae), a candidate for biological control of tree-of-heaven, Ailanthus altissima.

Ailanthus altissima (Mill.) Swingle, tree-of-heaven, is an invasive species native to Asia. It first was introduced into the United States in the 1700 s and now is distributed throughout much of North America. Mechanical and chemical controls are current suppression tactics, however, implementation is costly. A weevil, Eucryptorrhynchus brandti (Harold), was identified in China and imported for quarantine testing in 2004 as a potential biological control agent. Host specificity tests on adult feeding, larval development, and oviposition of this weevil were conducted from 2007 to 2011 on A. altissima and 29 nontarget species. Eucryptorrhynchus brandti adults fed significantly more on A. altissima foliage when compared with all test species. Range of means for feeding on A. altissima was 32.5-106.5 mm(2)/adult/d. In no-choice tests, Simarouba glauca DC, Leitneria floridana Chapm., and Citrus limon (L.) Burm. F., had feeding rates of only 10, 49, and 10%, respectively, compared with the level of feeding on A. altissima. The mean range of adult feeding by E. brandti on all other test species was <7% of feeding on A. altissima (0.0-3.3 ± 5.0 mm(2)/adult/d). In the no-choice larval inoculation tests, larval development only occurred in two of 10 L. floridana seedlings compared with seven of 10 A. altissima seedlings. In the no-choice oviposition tests, oviposition and subsequent larval development did not occur in L. floridiana, whereas all seven A. altissima seedlings supported oviposition and subsequent larval development. The weevil did not appear to be a threat to L. floridana or any other nontarget species tested. Therefore, we conclude that Eucryptorrhynchus brandti is highly host specific to A. altissima.

Feasibility of single-voxel MRS measurement of apparent diffusion coefficient of water in breast tumors.

We report initial results with single voxel spectroscopy (SVS) using diffusion weighting and localization by adiabatic selective refocusing (LASER) in breast tumors to measure the apparent diffusion coefficient of water (ADCw). This is a quick (30 s) and relatively easy method to implement compared with image-based diffusion measurements, and is insensitive to lipid signal contamination. The ADCw and concentration of total choline containing compounds [tCho] were evaluated for associations with each other and final pathologic diagnosis in 25 subjects. The average (+/- SD) ADCw in benign and malignant lesions was 1.96 +/- 0.47 mm(2)/s and 1.26 +/- 0.29 x 10(-3) mm(2)/s, respectively, P< 0.001. Receiver operating characteristic curve analysis showed an area under the curve of 0.92. Analysis of the single voxel (SV) ADCw and [tCho] showed significant correlation with a R(2) of 0.56, P< 0.001. Compared with more commonly used image-based methods of measuring water ADC, SV-ADCw is faster, more robust, insensitive to fat, and potentially easier to implement on standard clinical systems.

Maternal fear of hypoglycemia in their children with insulin dependent diabetes mellitus.

A modified version of the Hypoglycemia Fear Survey (HFS) was used to measure fear of hypoglycemia among mothers of 46 pre-adolescent children with IDDM. Glycosylated hemoglobin (HbA1) levels were measured, and episodes of severe hypoglycemia over the previous year were recorded. The relationships between the child's age, disease duration, HbA1, history of hypoglycemia, and maternal distress concerning hypoglycemia and HFS-P scores were explored. Maternal HFS-P scores were also compared to those of 78 IDDM adults. There was no relationship between children's age, disease duration, number of severe hypoglycemic episodes and maternal HFS-P scores. However, mothers whose children had passed out with hypoglycemia had higher HFS-P scores. Correlations between mothers' responses concerning their children's history of hypoglycemia and HFS-P scores suggested that their degree of distress associated with events which occurred when their child was asleep, or in a social situation was related to their total HFS-P, and Behavior and Worry subscores. Mothers demonstrated significantly greater fear than did adult patients themselves. The HFS-P may be an appropriate instrument for examining the psychological impact of differing treatment regimens on families of children with IDDM.

The Diabetes Control and Complications Trial: new challenges for the school nurse.

Results from the recently concluded Diabetes Control and Complications Trial confirm previously held beliefs regarding the relationships between hyperglycemia and the development of retinopathy, nephropathy, and neuropathy in insulin-dependent diabetes mellitus. Intensive therapy, now recommended for all patients 13 years and over, requires diabetes management throughout the school day and the active participation of the school nurse as a member of the child's health care team.

Accuracy of blood glucose estimation by children with IDDM and their parents.

OBJECTIVE: To evaluate the accuracy of blood glucose symptom recognition and subjective blood glucose estimation in insulin-dependent diabetic (IDDM) children and their parents. RESEARCH DESIGN AND METHODS: Blood glucose estimation questionnaires were completed 4 times/day at home during routine activities. A sequential sample of 19 families, who attended a pediatric diabetes clinic, with IDDM children less than 12 yr old and IDDM duration of greater than or equal to 9 mo comprised the study. RESULTS: Error grid analysis showed that both children and parents demonstrated poor accuracy, making clinically significant errors as frequently as clinically accurate estimates. The most common error was the failure to detect extreme blood glucose levels, with a significant tendency to underestimate hyperglycemia. Children often reported hypoglycemia when blood glucose was hyperglycemic. Confidence in the ability to estimate blood glucose was unrelated to measured accuracy. CONCLUSIONS: IDDM children and their parents demonstrated a higher rate of blood glucose estimation errors than IDDM adolescents and adults in previous studies. Even in families who use self-monitoring of blood glucose frequently, self-reported ability to recognize symptoms and estimate blood glucose should be viewed with caution. Families with IDDM children need more education about errors in symptom recognition and blood glucose estimation. They should also be encouraged to use self-monitoring of blood glucose before treating children's reported hypoglycemic symptoms whenever possible.

Stress in parents of children with diabetes mellitus.

The level of stress experienced in the parenting role by mothers of 49 children with insulin-dependent diabetes mellitus (IDDM) and its relationship to glycemic control was examined with the parenting stress index (PSI). A subsample of the research group of 25 children with diabetes (less than or equal to 11 yr old) was compared with an age-matched control group (n = 21) drawn from the original study of the PSI on total stress, parent- and child-domain, and subdomain scale scores. The two groups differed on one child-domain subscale, whereby children with diabetes are perceived by their mothers as more demanding than healthy controls. Three parent-subscale differences existed between the two groups, with mothers of children with diabetes reporting less attachment to their children, less spousal support, and poorer health. Analysis of the diabetes sample demonstrated significant stress on several of the child- and parent-domain subscales in a large proportion of the sample. Stress, at levels greater than or equal to 70th percentile of the control group, existed on the child scales of acceptability, mood, demanding behavior, and reinforcement for 51% of children with diabetes. Elevations associated with stress in the parenting role were evident on the scales associated with parental attachment, depression, and competence for 33% of parents. No differences in the level of glycosylated hemoglobin (HbA1) existed between children whose mothers reported high levels of stress in themselves and their children and those whose mothers reported little stress. Hierarchal regression analysis demonstrated a significant relationship between the child stress scale of distractibility, the use of self-monitoring blood glucose assessment, and low levels of HbA1.

Outpatient pediatric diabetes--I. Current practices.

A survey of pediatric diabetologists in the U.S. was made in an attempt to define current outpatient practices in diabetes subspecialty clinics. Survey questions addressed clinic organization, health care team members, content of histories and physical examinations, use of laboratory studies, patient education, therapeutic recommendations, self-management practices and screening procedures used to identify early diabetes-related complications. The results of the survey suggested similar clinic organization and operation in most settings; a high degree of reliance on glycosylated hemoglobin determinations; a preference for the use of NPH insulin; and a lack of credence given urinary glucose determinations. Additionally, screening tests for the development of complications are not performed with regularity.