Genetic diversity and cultivar variants in the NCGR cranberry (Vaccinium macrocarpon Aiton) collection.

The American cranberry (Vaccinium macrocarpon) is an endemic domesticated species that has become an economically important commercial fruit crop. The USDA-ARS National Clonal Germplasm Repository (NCGR) houses the national Vaccinium collection, which includes representatives of historical cranberry cultivars and wild-selected germplasm. The objective of this study wasto examine the genotypes of 271 cranberry plants from 77 accessions representing 66 named cultivars using 12 simple-sequence repeats to assess clonal purity and cultivar relatedness. Using principal components analysis and neighbour-joining based on estimated genetic distances between individuals, we identified 64 unique genotypes and observed that intracultivar variants (i.e. subclones) existed in the germplasm collection and in the commercial bogs where some accessions originated. Finally, through a comparison of the genotypes of this study with the previous studies, pedigree analysis and the study of the geographic distribution of cranberry diversity, we identified consensus genotypes for many accessions and cultivars. We highlight the important role that the NCGR collection playsfor ex situ conservation of cranberry germplasm for future breeders and researchers. The NCGR continues to search for historically relevant cultivars absent from the collection in an effort to preserve these genotypes before they are lost and no longer commercially grown.

Mining and validation of pyrosequenced simple sequence repeats (SSRs) from American cranberry (Vaccinium macrocarpon Ait.).

The American cranberry (Vaccinium macrocarpon Ait.) is a major commercial fruit crop in North America, but limited genetic resources have been developed for the species. Furthermore, the paucity of codominant DNA markers has hampered the advance of genetic research in cranberry and the Ericaceae family in general. Therefore, we used Roche 454 sequencing technology to perform low-coverage whole genome shotgun sequencing of the cranberry cultivar 'HyRed'. After de novo assembly, the obtained sequence covered 266.3 Mb of the estimated 540-590 Mb in cranberry genome. A total of 107,244 SSR loci were detected with an overall density across the genome of 403 SSR/Mb. The AG repeat was the most frequent motif in cranberry accounting for 35% of all SSRs and together with AAG and AAAT accounted for 46% of all loci discovered. To validate the SSR loci, we designed 96 primer-pairs using contig sequence data containing perfect SSR repeats, and studied the genetic diversity of 25 cranberry genotypes. We identified 48 polymorphic SSR loci with 2-15 alleles per locus for a total of 323 alleles in the 25 cranberry genotypes. Genetic clustering by principal coordinates and genetic structure analyzes confirmed the heterogeneous nature of cranberries. The parentage composition of several hybrid cultivars was evident from the structure analyzes. Whole genome shotgun 454 sequencing was a cost-effective and efficient way to identify numerous SSR repeats in the cranberry sequence for marker development.

Comparative Infectivity of Cronartium ribicola Aeciospores and Urediniospores in Genotypes of Ribes nigrum.

Hosts for the fungus Cronartium ribicola, causal agent of white pine blister rust (WPBR), include five-needle pines as aecial hosts, and currants and gooseberries as uredinial/telial hosts. Aeciospores produced on diseased pine, and urediniospores produced on diseased Ribes plants, can infect Ribes foliage. Resistance and susceptibility for both spore types have been reported for Ribes; however, the comparative infectivity of these spore types on clonal Ribes nigrum genotypes is under-described. Immunity, resistance, and susceptibility to WPBR resides at a clonal level in Ribes. Previous studies have emphasized fungal diversity or forestry considerations, rather than horticultural aspects. The objective of this study was to determine if aeciospores and urediniospores were equally infective to specific genotypes of black currant, Ribes nigrum, with differential responses. A family of 51 black currant genotypes from a cross between a known-immune cultivar containing the Cr gene and a susceptible cultivar was examined. Single-leaf softwood cuttings of each of these genotypes were artificially inoculated with a spore solution and incubated in airtight plastic containers within a growth chamber. Inoculations were replicated three times for each spore type. Twenty-two of the F1 genotypes did not develop uredia after artificial inoculation. These may be immune; 22 developed uredia after exposure to both types of inoculum, and were susceptible; whereas seven exhibited differential responses and may have some mechanism for resistance to WPBR other than the Cr gene. The infectivity of aeciospores and urediniospores was not significantly different on specific Ribes genotypes. Either spore type can therefore be considered equally effective as inoculum when screening for WPBR resistance or immunity in horticultural settings.

Screened hybrid density functionals applied to solids.

Hybrid Fock exchange/density functional theory functionals have shown to be very successful in describing a wide range of molecular properties. For periodic systems, however, the long-range nature of the Fock exchange interaction and the resultant large computational requirements present a major drawback. This is especially true for metallic systems, which require a dense Brillouin zone sampling. Recently, a new hybrid functional [HSE03, J. Heyd, G. E. Scuseria, and M. Ernzerhof, J. Chem. Phys. 118, 8207 (2003)] that addresses this problem within the context of methods that evaluate the Fock exchange in real space was introduced. We discuss the advantages the HSE03 functional brings to methods that rely on a reciprocal space description of the Fock exchange interaction, e.g., all methods that use plane wave basis sets. Furthermore, we present a detailed comparison of the performance of the HSE03 and PBE0 functionals for a set of archetypical solid state systems by calculating lattice parameters, bulk moduli, heats of formation, and band gaps. The results indicate that the hybrid functionals indeed often improve the description of these properties, but in several cases the results are not yet on par with standard gradient corrected functionals. This concerns in particular metallic systems for which the bandwidth and exchange splitting are seriously overestimated.

The incidence of emergence agitation associated with desflurane anesthesia in children is reduced by fentanyl.

UNLABELLED: The rapid emergence and recovery from general anesthesia provided by desflurane is associated with a frequent incidence of emergence agitation in children. We sought to determine the mean effective dose of fentanyl that would significantly reduce the incidence of emergence agitation while preserving rapid recovery. Thirty-two children undergoing adenoidectomy received general anesthesia with desflurane and a dose of fentanyl (1.25, 1.87, 2.8, and 4.2 microg/kg) determined by the classic up-down method. Recovery characteristics, including time to extubation, recovery, hospital discharge, agitation, pain, and vomiting, were recorded. Demographics and recovery features were assessed by analysis of variance and Kruskal-Wallis tests. The mean effective dose of fentanyl to reduce agitation was calculated with the Dixon-Massey method to be 2.5 +/- 6.2 microg. There were no significant differences when treatment groups were compared for recovery criteria. Postoperative emesis occurred in 75% of patients. The results of this study demonstrate that a dose of 2.5 microg/kg of fentanyl is sufficient to prevent emergence agitation while preserving the rapid recovery associated with desflurane anesthesia in children undergoing adenoidectomy. IMPLICATIONS: A dose of 2.5 microg/kg of fentanyl prevents emergence agitation associated with desflurane anesthesia in children undergoing adenoidectomy without delaying emergence.

The effect of intranasal fentanyl on the emergence characteristics after sevoflurane anesthesia in children undergoing surgery for bilateral myringotomy tube placement.

UNLABELLED: Children undergoing placement of bilateral myringotomy tubes (BMT) often exhibit pain-related behavior (agitation) in the postanesthesia care unit. We compared the emergence and recovery profiles of pediatric patients who received sevoflurane with or without supplementary intranasal fentanyl for BMT surgery. By using a prospective, double-blinded design, 150 children 6 mo to 5 yr of age, scheduled for routine BMT surgery, were anesthetized with sevoflurane (2%-3%) in a 60% N(2)O/O(2) gas mixture. Patients were randomized to receive equal volumes of intranasal saline (Control), 1 microg/kg fentanyl or 2 microg/kg fentanyl. A blinded observer evaluated each patient using a previously described 4-point agitation scale and the Steward recovery scale. Response to parental presence was observed after a score of six (full recovery) was achieved on the Steward recovery scale. There were no significant differences among the three groups regarding age, weight, surgeon, duration of anesthesia, or ear condition. Recovery times and emergence characteristic scores were not statistically different. Agitation scores were significantly reduced in the 2-microg/kg Fentanyl group as compared with the Control group (P = 0.012). Fentanyl 2 microg/kg is recommended to reduce the incidence of agitation seen in these patients. IMPLICATIONS: We examined the use of nasally administered fentanyl for the relief of agitation or discomfort after placement of bilateral myringotomy tubes in 150 children ages 6 mo to 5 yr using a prospective, double-blinded design. Fentanyl 2 microg/kg was found to reduce the incidence of agitation in these patients.

Formation of 8-iso-PGF(2alpha) and thromboxane A(2) by stimulation with several activators of phospholipase A(2) in the isolated human umbilical vein.

We investigated the effects of the phospholipase A(2) (PLA(2)) activators calcium ionophore A 23187, hydrogen peroxide (H(2)O(2)), bradykinin (BK), histamine and noradrenaline (NA) on the 8-iso-prostaglandin (PG)F(2alpha) formation in the isolated human umbilical vein and the isolated rabbit ear. For comparison, the influence of these substances on the thromboxane A(2) (TXA(2)) release was also investigated. The release of total (esterified as well as free) 8-iso-PGF(2alpha), free 8-iso-PGF(2alpha) and TXB(2), the stable metabolite of TXA(2), was determined by specific enzyme immunoassays. The results show that bolus injections of 5.4 mmol H(2)O(2), 30 nmol A 23187, 10 nmol BK, 50 nmol histamine and 20 nmol NA caused an increased release of total 8-iso-PGF(2alpha) in the umbilical vein and the rabbit ear. A perfusion with H(2)O(2) at a final concentration of 0.3 mM also increased the release of this isoprostane. Increased formation of free 8-iso-PGF(2alpha) was induced by A 23187 injection and by both modes of H(2)O(2) administration, but not by the other treatments. Bolus injections of A 23187, BK and histamine induced an increased release of TXB(2) in both organs. Both modes of H(2)O(2) administration and NA showed no releasing effects. In conclusion, our results show that the substances used are able to stimulate the formation of 8-iso-PGF(2alpha) concurrently with the release of PGs. This effect might be of pathophysiological relevance in inflammatory and cardiovascular diseases in which an enhanced release of free radicals, BK, histamine or NA play an important role.

Intramuscular rocuronium in infants and children: a multicenter study to evaluate tracheal intubating conditions, onset, and duration of action.

BACKGROUND: This multicenter, assessor-blinded, randomized study was done to confirm and extend a pilot study showing that intramuscular rocuronium can provide adequate tracheal intubating conditions in infants (2.5 min) and children (3 min) during halothane anesthesia. METHODS: Thirty-eight infants (age range, 3-12 months) and 38 children (age range, 1 to 5 yr) classified as American Society of Anesthesiologists physical status 1 and 2 were evaluated at four investigational sites. Anesthesia was maintained with halothane and oxygen (1% end-tidal concentration if <2.5 yr; 0.80% end-tidal concentration if >2.5 yr) for 5 min. One half of the patients received 0.45 mg/kg intravenous rocuronium. The others received 1 mg/kg (infants) or 1.8 mg/kg (children) of intramuscular rocuronium into the deltoid muscle. Intubating conditions and mechanomyographic responses to ulnar nerve stimulation were assessed. RESULTS: The conditions for tracheal intubation at 2.5 and 3 min in infants and children, respectively, were inadequate in a high percentage of patients in the intramuscular group. Nine of 16 infants and 10 of 17 children had adequate or better intubating conditions at 3.5 and 4 min, respectively, after intramuscular rocuronium. Better-than-adequate intubating conditions were achieved in 14 of 15 infants and 16 of 17 children given intravenous rocuronium. Intramuscular rocuronium provided > or =98% blockade in 7.4+/-3.4 min (in infants) and 8+/-6.3 min (in children). Twenty-five percent recovery occurred in 79+/-26 min (in infants) and in 86+/-22 min (in children). CONCLUSIONS: Intramuscular rocuronium, in the doses and conditions tested, does not consistently provide satisfactory tracheal intubating conditions in infants and children and is not an adequate alternative to intramuscular succinylcholine when rapid intubation is necessary.

Feasibility study for the measurement of a large set of triplet phases from a small protein.

The feasibility of measuring a set of triplet phases large enough to solve the structure of a small protein has been evaluated. A total of about 850 triplet phases have been measured from the tetragonal form of hen egg-white lysozyme. From these triplet phases, about 750 single phases can be derived. The experimental details of these measurements as well as the results, the values of the measured triplet phases, are reported. Additional experimental data from other small proteins are also presented.

A feasibility study of experimental triplet-phase determination in small proteins.

It is shown for the first time that the measurement of triplet phases of three-beam cases where strong structure factors are involved is possible in small protein structures, for example myoglobin. The exploitation of the triplet phase from the interference profile is not affected as long as the structure factors of unavoidable overlapping three-beam cases are small.

[Chronic hemorrhagic pancreatitis in gallbladder polyposis as an initial symptom of metachromatic leukodystrophy]. / Chronisch hämorrhagische Pankreatitis bei Gallenblasenpolyposis als Erstsymptom der metachromatischen Leukodystrophie.

A 4 1/2 year old boy without previous neurologic disorders developed chronic hemorrhagic pancreatitis and was shown to have polyposis of the gallbladder. Neurologic symptoms emerged at the age of 5 years. The sonographic pattern of an echogenic gallbladder was suspect of metachromatic leukodystrophy. The definitive diagnosis was made by the findings of very low arylsulfatase A activity in the white blood cells and deposits of sulfatides in the stroma of the polyps of the gallbladder.

Management of diabetes mellitus in children younger than 5 years of age.

Optimal treatment for children younger than 5 years of age with insulin-dependent diabetes mellitus is not well defined. Nineteen young children with this disease were treated with a program in which frequent home blood-glucose monitoring was used as the basis for an educational program emphasizing parental adjustment of insulin in response to current glucose levels and anticipated diet and exercise. Eleven children were treated from diagnosis (group I) and another eight (group D) were referred after less intensive treatment. The mean duration of observation of group I children was 13.6 months (range, six to 24 months). For group D, the mean time between diagnosis and referral was 14.9 months (range, seven to 24 months) and 14.6 months (range, six to 24 months) after referral. Before referral, there were 11 hospitalizations in group D. During the intensified program there were two hospitalizations in group D and one in group I. There were 3.3 episodes of severe hypoglycemia per child per 18 months in group D before referral, 1.7 episodes after referral, and 0.4 episodes in group I. Ten of 14 severe hypoglycemic episodes during intensified treatment occurred when there was no or infrequent home blood-glucose monitoring. Only four episodes seemed to have been unpredictable and unpreventable. Mean glycosylated hemoglobin levels were higher in group D patients when compared with both the duration of insulin-dependent diabetes mellitus and the time of initiation of intensified treatment. Mean daily insulin doses increased progressively in group I patients following diagnosis, and were comparable with those in group D patients at 15 and 18 months' duration of illness. Thus, frequency of hospitalization and severe hypoglycemia can be decreased in young children. Frequent home blood-glucose monitoring is required and extensive educational and psychosocial support is necessary for families to implement this intensive approach. The long-term effects on psychoneurological development need evaluation.