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Concussion is a complex, heterogeneous process affecting the brain. Accurate assessment and diagnosis and appropriate management of concussion are essential to ensure that athletes do not prematurely return to play or others to work or active military duty, risking re-injury. To date, clinical diagnosis relies primarily on evaluating subjects for functional impairment using instruments that include neurocognitive testing, subjective symptom report, and neurobehavioral assessments, such as balance and vestibular-ocular reflex testing. Structural biomarkers, defined as advanced neuroimaging techniques and biomarkers assessing neurotoxicity and immunoexcitotoxicity, may complement the use of functional biomarkers. We hypothesize that neurotoxicity AMPA, NMDA, and kainite receptor biomarkers might be utilized as a part of comprehensive approach to concussion evaluations, with the goal of increasing diagnostic accuracy and facilitating treatment planning and prognostic assessment.

As α-carboxy nucleoside phosphonates (α-CNPs) have demonstrated a novel mode of action of HIV-1 reverse transcriptase inhibition, structurally related derivatives were synthesized, namely the malonate 2, the unsaturated and saturated bisphosphonates 3 and 4, respectively and the amide 5. These compounds were evaluated for inhibition of HIV-1 reverse transcriptase in cell-free assays. The importance of the α-carboxy phosphonoacetic acid moiety for achieving reverse transcriptase inhibition, without the need for prior phosphorylation, was confirmed. The malonate derivative 2 was less active by two orders of magnitude than the original α-CNPs, while displaying the same pattern of kinetic behavior; interestingly the activity resides in the “L”-enantiomer of 2, as seen with the earlier series of α-CNPs. A crystal structure with an RT/DNA complex at 2.95 A resolution revealed the binding of the “L”-enantiomer of 2, at the polymerase active site with a weaker metal ion chelation environment compared to 1a (T-α-CNP) which may explain the lower inhibitory activity of 2.

The histone methyltransferase WHSC1 (also known as MMSET) is overexpressed in multiple myeloma (MM) as a result of the t(4;14) chromosomal translocation and in a broad variety of other cancers by unclear mechanisms. Overexpression of WHSC1 did not transform wild-type or tumor-prone primary hematopoietic cells. We found that ACA11, an orphan box H/ACA class small nucleolar RNA (snoRNA) encoded within an intron of WHSC1, was highly expressed in t(4;14)-positive MM and other cancers. ACA11 localized to nucleoli and bound what we believe to be a novel small nuclear ribonucleoprotein (snRNP) complex composed of several proteins involved in postsplicing intron complexes. RNA targets of this uncharacterized snRNP included snoRNA intermediates hosted within ribosomal protein (RP) genes, and an RP gene signature was strongly associated with t(4;14) in patients with MM. Expression of ACA11 was sufficient to downregulate RP genes and other snoRNAs implicated in the control of oxidative stress. ACA11 suppressed oxidative stress, afforded resistance to chemotherapy, and increased the proliferation of MM cells, demonstrating that ACA11 is a critical target of the t(4;14) translocation in MM and suggesting an oncogenic role in other cancers as well.

This study is part of a programmatic line of research into the use of basic positive reinforcement procedures for improving working memory in children with autism spectrum disorders. The authors evaluated the effects of multiple exemplar training, utilizing positive reinforcement, on performance of a “digit span backwards” task—a test of working memory that entails sequential relational responding. All three participants showed improved performance on directly trained stimuli as well as maintenance and generalization to untrained stimuli. The results provide further support for the general hypothesis that performance on working memory tasks is amenable to improvement via behavioral intervention and has implications for treating such tasks as relational operants. Implications for future research and the development of clinical interventions are discussed.

Birds employ numerous strategies to cope with seasonal fluctuations in high-quality habitat availability. Long distance migration is a common tactic; however, partial migration is especially common among broadly distributed species. Under partial migration systems, a portion of a species migrates, whereas the remainder inhabits breeding grounds year round. In this study, we identified effects of migratory behavior variation on genetic structure and diversity of American Kestrels (Falco sparverius), a widespread partial migrant in North America. American Kestrels generally migrate; however, a resident group inhabits the southeastern United States year round. The southeastern group is designated as a separate subspecies (F. s. paulus) from the migratory group (F. s. sparverius). Using mitochondrial DNA and microsatellites from 183 and 211 individuals, respectively, we illustrate that genetic structure is stronger among nonmigratory populations, with differentiation measures ranging from 0.060 to 0.189 depending on genetic marker and analysis approach. In contrast, measures from western North American populations ranged from 0 to 0.032. These findings suggest that seasonal migratory behavior is also associated with natal and breeding dispersal tendencies. We likewise detected significantly lower genetic diversity within nonmigratory populations, reflecting the greater influence of genetic drift in small populations. We identified the signal of population expansion among nonmigratory populations, consistent with the recent establishment of higher latitude breeding locations following Pleistocene glacial retreat. Differentiation of F. s. paulus and F. s. sparverius reflected subtle differences in allele frequencies. Because migratory behavior can evolve quickly, our analyses suggest recent origins of migratory American Kestrel populations in North America.

The Yellow Rail (Coturnicops noveboracencis) is among the most enigmatic and least studied North American birds. Nesting exclusively in marshes and wetlands, it breeds largely east of the Rocky Mountains in the northern United States and Canada, but there is an isolated population in southern Oregon once believed extirpated. The degree of connectivity of the Oregon population with the main population is unknown. We used mitochondrial DNA sequences (mtDNA) and six microsatellite loci to characterize the Yellow Rail's genetic structure and diversity patterns in six areas. Our mtDNA-based analyses of genetic structure identified significant population differentiation, but pairwise comparison of regions identified no clear geographic trends. In contrast, microsatellites suggested subtle genetic structure differentiating the Oregon population from those in the five regions sampled in the Yellow Rail's main breeding range. The genetic diversity of the Oregon population was also the lowest of the six r...

The Emergency Unemployment Compensation program created in the summer of 2008 provided for unprecedented extensions in the duration of unemployment insurance (UI) benefits. Combined with persistent high unemployment and historically long durations of unemployment during the 2008 and 2009 recession, this extension of UI has prompted renewed interest in the impact of UI benefits on job search, the duration of unemployment, and the unemployment rate. In a recent op-ed for the Wall Street Journal, Robert Barro contributed to this discussion by comparing the duration of unemployment observed in the current recession with that of 1982 and estimating how much lower the unemployment rate would have been in the summer of 2010 if it had not been for the extension of the duration of UI benefits. His conclusion, that the unemployment rate would have been 2.7 points lower in the absence of UI extensions, is based on a questionable assumption—that the severity of the 2008-09 recession could not be responsible. This paper uses multiple regression analysis to estimate the impact of extended UI benefits on the unemployment rate after controlling for the severity of the recent recession. The extension of UI is found to have a positive and significant impact on the national unemployment rate, but the impact is not as large as Barro suggests. The UI benefit extensions that have occurred between the summer of 2008 and the end of 2010 are estimated to have had a cumulative effect of raising the unemployment rate by .77 to 1.54 percentage points.

It has been proposed that people with autism spectrum disorder (ASD) have abnormal morphometry and development of the amygdala and hippocampus (AH). However, previous reports are inconsistent, perhaps because they included people of different ASD diagnoses, ages, and health. We compared, using magnetic resonance imaging, the in vivo anatomy of the AH in 32 healthy individuals with Asperger syndrome (12-47 years) and 32 healthy controls who did not differ significantly in age or IQ. We measured bulk (gray + white matter) volume of the AH using manual tracing (MEASURE). We first compared the volume of AH between individuals with Asperger syndrome and controls and then investigated age-related differences. We compared differences in anatomy before, and after, correcting for whole brain size. There was no significant between group differences in whole brain volume. However, individuals with Asperger syndrome had a significantly larger raw bulk volume of total (P

Aims and methodTo identify the provision of psychosocial assessments for all people attending an accident and emergency department in Ireland with a presentation indicative of self-harm over 12 months and to investigate whether the National Institute for Health and Clinical Excellence (NICE) guidelines for self-harm were met.ResultsA total of 834 attendances for self-harm were recorded. A psychosocial assessment was undertaken by a member of the liaison psychiatry team in 59% of attendances. Single male patients under 45 years of age represented 39% of those who did not receive a psychosocial assessment.Clinical implicationsSingle men under the age of 45 years represent a vulnerable group in which levels of psychosocial assessment need to be optimised in order to meet the NICE guidelines for standards of care.

Piping Plover (Charadrius melodus) is a migratory shorebird that is listed as endangered in Canada and the U.S. Great Lakes and as threatened throughout the rest of its breeding and winter range. We undertook a comprehensive molecular-genetic investigation to (1) address subspecific taxonomy, (2) characterize population genetic structure, and (3) infer past bottlenecks and demographic processes in this species. Analyses included individuals from 23 U.S. states and Canadian provinces and were based on mitochondrial DNA sequences (580 base pairs, n = 245) and 8 nuclear microsatellite loci (n = 229). Our findings provide support for separate Atlantic and Interior subspecies (C. m. melodus and C. m. circumcinctus, respectively). Birds from the Great Lakes region were allied with the Interior subspecies and should be referred to as C. m. circumcinctus. Population genetic analyses illustrated stronger genetic structure among Atlantic than among Interior birds, which may reflect reduced natal- and bree...

A preventive intervention for reducing physical and relational aggression, peer victimization, and increasing prosocial behavior was developed for use in early childhood classrooms. Nine classrooms were randomly assigned to be intervention rooms ( N = 202 children) and nine classrooms were control rooms ( N = 201 children). Classroom was the unit of analysis and both observations and teacher-reports were obtained at pre and post-test. Focus groups were used to develop the initial program. The 6-week program consisted of developmentally appropriate puppet shows, active participatory sessions, passive concept activities and in vivo reinforcement periods. Preliminary findings suggest that the “Early Childhood Friendship Project” tended to reduce physical and relational aggression, as well as physical and relational victimization and tended to increase prosocial behavior more for intervention than control classrooms. Teachers and interventionists provided positive evaluations of the program and there is evidence for appropriate program implementation.

The homo-oxo-calix[3]arene-facilitated electrochemistry of dopamine at the polarised liquid|liquid interface is presented. Cyclic voltammetry indicated that the dopamine: homo-oxo-calix[3]arene complex formed at the interface between water and 1,2-dichloroethane had a 1:2 stoichiometry, which was confirmed by 1 H NMR titration. The electrochemically-determined association constants for the homo-oxo-calix[3]arene complexes with dopamine (log β 8.3), Na + (8.0) and K + (7.5) showed that the ionophore was more selective for dopamine over the two alkali metal cations than the dibenzo-18-crown-6 ionophore. Square wave voltammetry was employed for assessment of low concentration measurements, with a limit of detection of 3.8 μM dopamine achieved. These results show that oxo-calixarene-based ionophores are useful reagents for the detection of dopamine at the interface between two immiscible electrolyte solutions.

Population genetics plays an increasingly important role in the conservation and management of declining species, particularly for defining taxonomic units. Subspecies are recognized by several conservation organizations and countries and receive legal protection under the US Endangered Species Act (ESA). Two subspecies of spotted owls, northern (Strix occidentalis caurina) and Mexican (S. o. lucida) spotted owls, are ESA-listed as threatened, but the California (S. o. occidentalis) spotted owl is not listed. Thus, determining the boundaries of these subspecies is critical for effective enforcement of the ESA. We tested the validity of previously recognized spotted owl subspecies by analysing 394 spotted owls at 10 microsatellite loci. We also tested whether northern and California spotted owls hybridize as suggested by previous mitochondrial DNA studies. Our results supported current recognition of three subspecies. We also found bi-directional hybridization and dispersal between northern and California spotted owls centered in southern Oregon and northern California. Surprisingly, we also detected introgression of Mexican spotted owls into the range of northern spotted owls, primarily in the northern part of the subspecies’ range in Washington, indicating long-distance dispersal of Mexican spotted owls. We conclude with a discussion of the conservation implications of our study.

Acetamidobenzamide 1 exists in two crystal polymorphic forms: α and β. The α form features an approximately planar molecular conformation and an intramolecular N−H...O hydrogen bond. In the β form, the amido groups are out of the plane of the benzene ring and no intramolecular hydrogen bonds are present. In our study, the α form was found to be the more readily obtainable initially, usually as well-formed needles. 2-Acetamido-4-chlorobenzamide 8 and 2-acetamido-5- chlorobenzamide 9 were prepared and assessed as additives in recrystallizations of 2- acetamidobenzamide. In the presence of compound 8, attempted recrystallization of 2- acetamidobenzamide gave crystals of the anhydrous form of 2-methylquinazol-4-one 4. Addition of compound 9 had no effect on the crystal form obtained. 10,11-Dihydro-5-acetyl- dibenzo(b,e)(1,4)diazepin-11-one 14 was prepared as a conformational mimic of the molecular conformation of the β form of 2-acetamidobenzamide. Compound 14 had no effect on the recrystallization of 2-acetamidobenzamide from acetone when added in 10% w/w or from water when added in 1% w/w. When 14 was added in 10% w/w quantities to crystallizations of 2- acetamidobenzamide from water, fine crystallites were obtained which gave infra-red spectra consistent with the α form, but gave PXRD patterns suggesting the presence of another form or compound. Disruption of the intramolecular hydrogen bonds in the α form of 2- acetamidobenzamide 1 by direct trimethylsilylation gave mixtures of 1 and 2-methylquinazol-4- one 4. Indirect trimethylsilylation by reaction of 2-methyl-4H-benzo(d)(1,3)oxazin-4-one 3 with tris(trimethylsilyl)amine gave crystals of the anhydrous form of 2-methylquinazol-4-one 4. 2- Phthalimidobenzoic acid 18 and 2-phthalimidobenzamide 19 were prepared as structural analogues of compound 1 which are incapable of possessing an intramolecular hydrogen bond. Addition of compound 18 to recrystallizations of 2-acetamidobenzamide 1 had no effect on the outcome. However addition of compound 19 resulted in needle-like crystals of the α form of considerably reduced dimensions.

In December 2001, Canada's response to the international political climate was launched by the creation of the Chemical, Biological, Radiological/Nuclear Research & Technology Initiative (CRTI). The National Biological Dosimetry Response Plan (NBDRP), established through partnering the expertise of three federal departments and one university, was created in response to this initiative. The NBDRP objectives were to develop a network of laboratories with expertise to perform biological dosimetry by cytogenetics and to investigate new technologies that may be applicable in the development of the new biodosimetry program. Since the creation of the NBDRP, Canada has made significant progress in enhancing expertise and resources to be better prepared for radiological/nuclear events. Through participation in exercises, the existing capacities were tested and recommendations for improvements were made. This paper describes the results from two exercises. The first exercise was designed to test the culturing, analysis, and reporting procedures within a single laboratory, and the second exercise was intended to test the capacity of the NBDRP. Future exercises will further challenge the network resulting in an improved national response capability.

While biodosimetry is a valuable tool in radiation dose assessment, the dicentric assay, which is the most validated method to date, requires some degree of technical competence. Recently published ISO guidelines indicate the need for documenting competence and establishment of quality control programs. Inter-laboratory comparisons are required to document the ability to perform reproducible and accurate assessments. FOI and DRDC Ottawa have conducted an initial limited biodosimetry exercise inter-comparison for quality assurance purposes. The exercise involved blinded exchange of three previously prepared slides from each laboratory from samples that had been evaluated for each lab's dose–response curve. Approximately 100 cells from each slide were evaluated and aberration frequencies reported and compared to the expected frequencies. The limited number of cells evaluated for each sample could not permit statistically distinguishing a 20% difference in all the samples. However, the results indicated reasonable agreement in analyses for all samples for triage purposes. Comparison of aberration frequencies, rather than dose estimates, further illustrates consistent scoring criteria between the two laboratories. The exercise conducted by FOI and DRDC Ottawa provided an efficient means of documenting expertise. Such cooperation further establishes the international biodosimetry network and ensures our readiness for emergency response.

We examined the genetic structure of snowy plovers (Charadrius alexandrinus) in North America, the Caribbean, and the west coast of South America to quantify variation within and among breeding areas and to test the validity of three previously recognized subspecies. Sequences (676 bp) from domains I and II of the mitochondrial control region were analyzed for 166 snowy plovers from 20 breeding areas. Variation was also examined at 10 microsatellite loci for 144 snowy plovers from 14 breeding areas. The mtDNA and microsatellite data provided strong evidence that the Puerto Rican breeding group is genetically divergent from sites in the continental U.S. (net sequence divergence = 0.38%; F ST for microsatellites = 0.190). Our data also revealed high levels of differentiation between sites from South America and North America (net sequence divergence = 0.81%; F ST for microsatellites = 0.253). In contrast, there was little genetic structure among breeding sites within the continental U.S. Our results suggest that snowy plovers in Florida should be considered part of C. a. nivosus (rather than part of C. a. tenuirostris, where they are currently placed), whereas snowy plovers from Puerto Rico should be considered part of C. a. tenuirostris. Snowy plovers in South America should remain a separate subspecies (C. a. occidentalis). Although U.S. Pacific and Gulf Coast breeding areas were not genetically distinct from other continental U.S. sites, demographic isolation, unique coastal habitats, and recent population declines suggest they warrant special concern.

The U.S. Endangered Species Act (ESA) allows listing of subspecies and other groupings below the rank of species. This provides the U.S. Fish and Wildlife Service and the National Marine Fisheries Service with a means to target the most critical unit in need of conservation. Although roughly one-quarter of listed taxa are subspecies, these management agencies are hindered by uncertainties about taxonomic standards during listing or delisting activities. In a review of taxonomic publications and societies, we found few subspecies lists and none that stated standardized criteria for determining subspecific taxa. Lack of criteria is attributed to a centuries-old debate over species and subspecies concepts. Nevertheless, the critical need to resolve this debate for ESA listings led us to propose that minimal biological criteria to define disjunct subspecies (legally or taxonomically) should include the discreteness and significance criteria of distinct population segments (as defined under the ESA). Our subspecies criteria are in stark contrast to that proposed by supporters of the phylogenetic species concept and provide a clear distinction between species and subspecies. Efforts to eliminate or reduce ambiguity associated with subspecies-level classifications will assist with ESA listing decisions. Thus, we urge professional taxonomic societies to publish and periodically update peer-reviewed species and subspecies lists. This effort must be paralleled throughout the world for efficient taxonomic conservation to take place.

Taxonomic relationships among red tree voles (Phenacomys longicaudus longicaudus, P. l. silvicola), the Sonoma tree vole (P. pomo), the white-footed vole (P. albipes), and the heather vole (P. intermedius) were examined using 664 base pairs of the mitochondrial cytochrome b gene. Results indicate specific differences among red tree voles, Sonoma tree voles, white-footed voles, and heather voles, but no clear difference between the 2 Oregon subspecies of red tree voles (P. l. longicaudus and P. l. silvicola). Our data further indicated a close relationship between tree voles and albipes, validating inclusion of albipes in the subgenus Arborimus. These 3 congeners shared a closer relationship to P. intermedius than to other arvicolids. A moderate association between pomo and albipes was indicated by maximum parsimony and neighbor-joining phylogenetic analyses. Molecular clock estimates suggest a Pleistocene radiation of the Arborimus clade, which is concordant with pulses of diversification observe...

Pre-cooled bulbs of two dutch iris (Iris ×hollandica) cultivars, Ideal and White Wedgewood, were grown and harvested as cut flowers in four production systems in a tobacco (Nicotiana tabacum) transplant greenhouse from late October until late January in two consecutive production years (2000-01 and 2001-02). All production systems (lily crates, lay-flat bags, pots, and float trays) utilized the same commercial peat-vermiculite, tobacco germination substrate. Stems developed more quickly but were shorter and lighter in 2001-02 than 2000-01 due to warmer growing conditions. Stems grown in float trays were shorter and lighter than other treatments in 2000-01 but similar to the others in 2001-02. Stems grown in lay-flat bags flowered earlier with similar or greater stem lengths and fresh weights as the other systems. Stems of `White Wedgewood' were longer and heavier than `Ideal'. In general, `White Wedgewood' provided more consistent production than `Ideal' in both production seasons. An economic analysis in this study concludes that a grower is unlikely to make money growing dutch iris in a tobacco transplant greenhouse using these production systems unless there is a targeted local market.

Recent population expansion of Barred Owls ( Strix varia) into western North America has led to concern that they may compete with and further harm the Northern Spotted Owl ( S. occidentalis caurina), which is already listed as threatened under the U.S. Endangered Species Act (ESA). Because they hybridize, there is a legal need under the ESA for forensic identification of both species and their hybrids. We used mitochondrial control-region DNA and amplified fragment-length polymorphism (AFLP) analyses to assess maternal and biparental gene flow in this hybridization process. Mitochondrial DNA sequences (524 base pairs) indicated large divergence between Barred and Spotted Owls (13.9%). Further, the species formed two distinct clades with no signs of previous introgression. Fourteen diagnostic AFLP bands also indicated extensive divergence between the species, including markers differentiating them. Principal coordinate analyses and assignment tests clearly supported this differentiation. We found that hybrids had unique genetic combinations, including AFLP markers from both parental species, and identified known hybrids as well as potential hybrids with unclear taxonomic status. Our analyses corroborated the findings of extensive field studies that most hybrids genetically sampled resulted from crosses between female Barred Owls and male Spotted Owls. These genetic markers make it possible to clearly identify these species as well as hybrids and can now be used for research, conservation, and law enforcement. Several legal avenues may facilitate future conservation of Spotted Owls and other ESA-listed species that hybridize, including the ESA similarity-of-appearance clause (section 4[e]) and the Migratory Bird Treaty Act. The Migratory Bird Treaty Act appears to be the most useful route at this time. Resumen: La reciente expansion de la poblacion de Strix varia hacia el oeste de Norte America ha llevado a la preocupacion de que puede competir con y posteriormente danar a S. occidentalis caurina, que esta enlistada como amenazada en el Acta de Especies en Peligro de E. U. (AEP). Debido a que hibridan, por mandato de AEP hay una necesidad legal de la identificacion forense de ambas especies y sus hibridos. Utilizamos analisis de ADN de la region de control mitocondrial y polimorfismo de longitud de fragmento amplificado (PLFA) para evaluar el flujo genico materno y biparental de este proceso de hibridacion. Las secuencias de ADN mitocondrial (524 pares de bases) indicaron una amplia divergencia ente Strix varia y S. occidentalis caurina (13.9%). Mas aun, las especies formaron dos clados diferentes sin signos de introgresion previa. Catorce bandas diagnostico de PLFA tambien indicaron divergencia extensiva entre especies, incluyendo marcadores que diferencian a las especies. Analisis de componentes principales y pruebas de acomodo claramente sustentaron a esta diferenciacion. Encontramos que los hibridos tenian combinaciones geneticas unicas, incluyendo marcadores PLFA de ambas especies parentales e identificaban a hibridos conocidos asi como a los potenciales hibridos con estatus taxonomico incierto. Nuestros analisis corroboraron extensivos estudios de campo que la mayoria de los hibridos muestreados geneticamente resultaron de cruzas entre hembras de Strix varia y machos de S. occidentalis caurina. Estos marcadores geneticos hacen que sea posible identificar claramente a estas especies asi como a sus hibridos y ahora pueden ser utilizados para investigacion, conservacion y aplicacion de la ley. Varias rutas legales pueden facilitar la conservacion futura de S. occidentalis caurina y otras especies enlistadas en AEP que hibridan, incluyendo la clausula de similitud de apariencia de AEP (seccion 4[e]) y el Acta del Tratado de Aves Migratorias. Por el momento, el Acta del Tratado de Aves Migratorias parece ser la ruta mas util.

Hierarchical genetic structure was examined in the three geographically-defined subspecies of spotted owl (Strix occidentalis) to define relationships among subspecies and quantify variation within and among regional and local populations. Sequences (522 bp) from domains I and II of the mitochondrial control region were analyzed for 213 individuals from 30 local breeding areas. Results confirmed significant differences between northern spotted owls and the other traditional geographically defined subspecies but did not provide support for subspecific level differences between California and Mexican spotted owls. Divergence times among subspecies estimated with a 936 bp portion of the cytochrome b gene dated Northern and California/Mexican spotted owl divergence time to 115,000–125,000 years ago, whereas California/Mexican spotted owl divergence was estimated at 15,000 years ago. Nested clade analyses indicated an association between California spotted owl and Mexican spotted owl haplotypes, implying historical contact between the two groups. Results also identified a number of individuals geographically classified as northern spotted owls (S. o. caurina) that contained haplotypes identified as California spotted owls (S. o. caurina). Among all northern spotted owls sampled (n=131), 12.9% contained California spotted owl haplotypes. In the Klamath region, which is the contact zone between the two subspecies, 20.3% (n=59) of owls were classified as California spotted owls. The Klamath region is a zone of hybridization and speciation for many other taxa as well. Analyses of population structure indicated gene flow among regions within geographically defined subspecies although there was significant differentiation among northern and southern regions of Mexican spotted owls. Among all areas examined, genetic diversity was not significantly reduced except in California spotted owls where the southern region consists of one haplotype. Our results indicate a stable contact zone between northern and California spotted owls, maintaining distinct subspecific haplotypes within their traditional ranges. This supports recovery efforts based on the traditional subspecies designation for the northern spotted owl. Further, although little variation was found between California and Mexican spotted owls, we suggest they should be managed separately because of current isolation between groups.

We used Illumina sequencing to develop eight novel microsatellite loci for Maasai giraffe (Giraffa camelopardalis tippelskirchi), and screened them using fecal DNA. Genetic diversity was assessed for 40 individuals from the Katavi National Park/Rukwa Game Reserve ecosystem in Tanzania. The number of alleles per locus ranged from 2 to 14 (mean = 6.2) and mean expected heterozygosity was moderate (range 0.025–0.838, mean = 0.527). These markers were successfully tested using degraded DNA and may be useful for future studies of giraffe, especially those that use non-invasive sampling techniques.

Drug shortages have increasingly been a problem for pharmacists and clinicians over the past decade. Clinicians need to be aware of the various causes of drug shortages and the issues that may arise as a result, particularly as they relate to medication safety. Numerous resources and strategies are available to mitigate the effects of drug shortages, and clinicians should work with their health care team and the patient to determine the best option when faced with a drug shortage that affects patient care.

To determine whether extremely low frequency electromagnetic fields can alter average free cytosolic calcium ion concentrations [Ca2+]i and transient increases in [Ca2+]i in populations of ROS 17/2.8 cells.Cells loaded with the calcium-selective luminescent photoprotein, aequorin, were placed in the bottom of a sample chamber, which was inserted into the gap of a previously described air gap reactor system where they were exposed either to sinusoidal magnetic fields at a variety of frequencies and flux densities or to sham conditions. Real-time recordings of photon counts due to aequorin luminescence were obtained and data were analysed with the use of probit plots.Probit plots of data obtained from cells exposed to the various magnetic fields were virtually superimposable over the data obtained for the same cultures during pre- and post-exposure sham or no-field periods.These experiments provided no evidence for any effects of ELF EMF, either positive or negative, on either average [Ca2+]i or on transient increases in [Ca2+]i.

We examined genetic variation, populationstructure, and definition of conservation unitsin Spotted Owls (Strix occidentalis). Spotted Owls are mostly non-migratory,long-lived, socially monogamous birds that havedecreased population viability due to theiroccupation of highly-fragmented latesuccessional forests in western North America. To investigate potential effects of habitatfragmentation on population structure, we usedrandom amplified polymorphic DNA (RAPD) toexamine genetic variation hierarchicallyamong local breeding areas, subregionalgroups, regional groups, and subspeciesvia sampling of 21 breeding areas (276individuals) among the three subspecies ofSpotted Owls. Data from 11 variable bandssuggest a significant relationship betweengeographic distance among local breeding groupsand genetic distance (Mantel r = 0.53, P< 0.02) although multi-dimensional scaling ofthree significant axes did not identifysignificant grouping at any hierarchical level. Similarly, neighbor-joining clustering ofManhattan distances indicated geographicstructure at all levels and identified MexicanSpotted Owls as a distinct clade. RAPDanalyses did not clearly differentiate NorthernSpotted Owls from California Spotted Owls. Among Northern Spotted Owls, estimates ofpopulation differentiation (FST) rangedfrom 0.27 among breeding areas to 0.11 amongregions. Concordantly, within-group agreementvalues estimated via multi-response permutationprocedures of Jaccard's distances ranged from0.22 among local sites to 0.11 among regions. Pairwise comparisons of FST andgeographic distance within regions suggestedonly the Klamath region was in equilibrium withrespect to gene flow and genetic drift. Merging nuclear data with recent mitochondrialdata provides support for designation of anEvolutionary Significant Unit for MexicanSpotted Owls and two overlapping ManagementUnits for Northern and California Spotted Owls.

We describe the isolation and characterization of eight microsatellite loci from the double-crested cormorant (Phalacrocorax auritus). Genetic variability was assessed using 60 individuals from three populations. All loci were variable with the number of alleles ranging from two to 17 per locus, and observed heterozygosity varying from 0.05 to 0.89. No loci showed signs of linkage disequilibrium and all loci conformed to Hardy–Weinberg equilibrium frequencies. Further, all loci amplified and were polymorphic in two related Phalacrocorax species. These loci should prove useful for population genetic studies of the double-crested cormorant and other pelecaniform species.

Multiple factors contribute to the development of posttraumatic empyema. These factors include the conditions under which the tube is inserted (emergent or urgent), the mechanism of injury, retained hemothorax, and ventilator care. The incidence of empyema in placebo groups ranges between 0 and 18%. The administration of antibiotics for longer than 24 hours did not seem to significantly reduce this risk compared with a shorter duration, although the numbers in each series were small. Most reports found a significant reduction in pneumonitis when patients received prolonged prophylactic antibiotics. This use of antibiotics might possibly be better described as presumptive therapy rather than prophylactic.

Fullen et al. first described the role for antibiotics in patients sustaining penetrating abdominal injuries. They retrospectively reviewed 295 patients who underwent celiotomy after sustaining penetrating abdominal wounds and categorized patients according to the timing of their first antibiotic dose: preoperative, n 5 16; intraoperative, n 5 98; and postoperative, n 5 81. The reported rate of trauma-related infections (incisional and intra-abdominal abscess) were 7%, 33%, and 30%, respectively. Individuals with colon injuries had postoperative infection rates of 11%, 57%, and 70% for each group, respectively. These rates remained constant even when the data were analyzed for additional risk factors, including the number of associated intra-abdominal organs injured, frequency of shock, and need for transfusion of blood products. The average time from hospital admission to laparotomy was the same for all three groups. Regardless of whether the observed difference was caused by the intraoperative or postoperative groups having a longer interval between injury and antibiotic administration or that the preoperative group had antibiotics circulating at the time of incision, this was the first study to suggest that the timing of antibiotic administration can impact the development of injury-related infections in patients with penetrating abdominal injuries. The importance of broad-spectrum antibiotic coverage for these patients was demonstrated by Thadepalli et al. in 1973. This study was a prospective, randomly assigned comparison of kanamycin and cephalothin to kanamycin and clindamycin. Both antibiotic combinations were administered preoperatively. The clindamycin group had a significantly lower rate of infection in the postoperative period compared with the cephalothin group (10% vs. 27%). They further demonstrated that the difference was caused by significantly more anaerobic infections in the cephalothin group (21%) compared with the clindamycin group (2%). These two studies demonstrated a significantly lower rate of infection when antibiotics providing aerobic and anaerobic coverage are administered before operative treatment. Prophylactic antibiotics for patients sustaining penetrating abdominal injuries with intestinal contamination have a role for reducing the rate of incisional wound infection subjected to gastrointestinal soiling. A single dose providing sufficient concentration within the wound during the vulnerable period is optimal. The other aspect of prophylactic antibiotic administration in trauma is the potential therapeutic role. The problem is to define the time period when contamination of the abdominal cavity becomes an established infection. At celiotomy, the intestinal wound is closed, eliminating further contamination and soiling of the peritoneal cavity. Thus, no further antibiotic should be necessary. Surgeons have concluded that “prophylactic antibiotics” in penetrating abdominal trauma can reduce the incidence of postoperative infectious complications. Since the mid-1970s, no study has included a placebo control group because of the high incidence of infectious complications after intestinal injury. However, many studies in the past 2 decades have compared various antibiotic regimens to evaluate single agents versus combination regimens, duration of administration, and, more recently, the pharmacokinetics and cost implications of single versus combination therapy.

The Arp2/3 complex, a stable assembly of two actin-related proteins (Arp2 and Arp3) with five other subunits, caps the pointed end of actin filaments and nucleates actin polymerization with low efficiency. WASp and Scar are two similar proteins that bind the p21 subunit of the Arp2/3 complex, but their effect on the nucleation activity of the complex was not known. We report that full-length, recombinant human Scar protein, as well as N-terminally truncated Scar proteins, enhance nucleation by the Arp2/3 complex. By themselves, these proteins either have no effect or inhibit actin polymerization. The actin monomer-binding W domain and the p21-binding A domain from the C terminus of Scar are both required to activate Arp2/3 complex. A proline-rich domain in the middle of Scar enhances the activity of the W and A domains. Preincubating Scar and Arp2/3 complex with actin filaments overcomes the initial lag in polymerization, suggesting that efficient nucleation by the Arp2/3 complex requires assembly on the side of a preexisting filament—a dendritic nucleation mechanism. The Arp2/3 complex with full-length Scar, Scar containing P, W, and A domains, or Scar containing W and A domains overcomes inhibition of nucleation by the actin monomer-binding protein profilin, giving active nucleation over a low background of spontaneous nucleation. These results show that Scar and, likely, related proteins, such as the Cdc42 targets WASp and N-WASp, are endogenous activators of actin polymerization by the Arp2/3 complex.

Purpose: We compare noncontrast enhanced computerized tomography (CT) and excretory urography (IVP) in the evaluation of acute flank pain.Materials and Methods: A total of 40 consecutive patients presenting to the emergency department with acute flank pain were evaluated with noncontrast CT, films of the kidneys, ureters and bladder, and IVP. The patients were treated according to the clinical picture. All 40 sets of evaluations were later assessed randomly by an independent consultant radiologist for the presence, size and location of a stone, ureteral dilatation and secondary signs of ureteral obstruction.Results: Of 40 patients 12 had no calculus and 28 had a calculus confirmed on removal or documented passage of a stone. Absence of a stone was based on clinical and radiological followup with clinical resolution. CT revealed all 28 calculi and no calculus in 11 of 12 patients with 100% sensitivity and 92% specificity. IVP demonstrated 18 calculi (64% sensitivity) and no calculus in 11 of 12 pat...

We identified four diagnostic microsatellite loci that distinguish spotted owls (Strix occidentalis), barred owls (Strix varia), F 1 hybrids and backcrosses. Thirty-four out of 52 loci tested (65.4%) successfully amplified, and four of these loci (11.8%) had allele sizes that did not overlap between spotted and barred owls. The probability of correctly identifying a backcross with these four loci is 0.875. Genotyping potential hybrid owls with these markers revealed that field identifications were often wrong. Given the difficulty of identifying hybrids in the field, these markers will be useful for hybrid identification, law enforcement and spotted owl conservation.

Identification of relationships among geographically distinct populations of migratory species can provide an understanding of breeding and natal philopatry, migration pathways, and population mixing during winter. We used random amplified polymorphic DNA (RAPD) analyses to search for markers specific to difficult-to-differentiate shorebird species (e.g. long-billed dowitcher Limnodromus scolopaceus and short-billed dowitcher L. griseus) as well as geographically distinct breeding populations of Hudsonian godwits Limosa haemastica, red-necked phalaropes Phalaropus lobatus, semipalmated plovers Charadrius semipalmatus, dunlin Calidris alpina, pectoral sandpipers C. melanotos, semipalmated sandpipers C. pusilla and western sandpipers C. mauri. Markers clearly differentiated all shorebird species. Estimates of population differentiation varied greatly among species (FST = 0.095‐0.685) and correlated with interspecific variation in philopatry and geographical separation of breeding populations. We assigned individuals to putative breeding locales with greater certainty in welldifferentiated species than in poorly differentiated species. Our findings indicate specific phylogeographical structure varies among species, which has strong implications for conservation of habitats within migratory corridors. We suggest that RAPDs are useful in identifying geographical populations of migratory species and that molecular markers should be considered for tracking migratory birds throughout the annual cycle.

SSR_pipeline is a flexible set of programs designed to efficiently identify simple sequence repeats (e.g., microsatellites) from paired-end high-throughput Illumina DNA sequencing data. The program suite contains 3 analysis modules along with a fourth control module that can automate analyses of large volumes of data. The modules are used to 1) identify the subset of paired-end sequences that pass Illumina quality standards, 2) align paired-end reads into a single composite DNA sequence, and 3) identify sequences that possess microsatellites (both simple and compound) conforming to user-specified parameters. The microsatellite search algorithm is extremely efficient, and we have used it to identify repeats with motifs from 2 to 25 bp in length. Each of the 3 analysis modules can also be used independently to provide greater flexibility or to work with FASTQ or FASTA files generated from other sequencing platforms (Roche 454, Ion Torrent, etc.). We demonstrate use of the program with data from the brine fly Ephydra packardi (Diptera: Ephydridae) and provide empirical timing benchmarks to illustrate program performance on a common desktop computer environment. We further show that the Illumina platform is capable of identifying large numbers of microsatellites, even when using unenriched sample libraries and a very small percentage of the sequencing capacity from a single DNA sequencing run. All modules from SSR_pipeline are implemented in the Python programming language and can therefore be used from nearly any computer operating system (Linux, Macintosh, and Windows).

Background: Down’s syndrome (DS) is the most common genetic cause of intellectual disability. People with DS are at an increased risk of Alzheimer’s disease (AD) compared to the general population. Neuroimaging studies of AD have focused on medial temporal structures; however, to our knowledge, no in vivo case–control study exists comparing the anatomy of dementia in DS to people with AD in the general population. We therefore compared the in vivo brain anatomy of people with DS and dementia (DS+) to those with AD in the general population. Method: Using MRI in 192 adults, we compared the volume of whole brain matter, lateral ventricles, temporal lobes and hippocampus in DS subjects with and without dementia (DS+, DS-), to each other and to three non-DS groups. These included one group of individuals with AD and two groups of controls (each age-matched for their respective DS and general population AD cohorts). Results: AD and DS+ subjects showed significant reductions in the volume of the whole brain, hippocampus and temporal lobes and a significant elevation in the volume of the lateral ventricle, compared to their non-demented counterparts. People with DS+ had a smaller reduction in temporal lobe volume compared to individuals with AD. Conclusions: DS+ and AD subjects have a significant reduction in volume of the same brain regions. We found preliminary evidence that DS individuals may be more sensitive to tissue loss than others and have less ‘cognitive reserve’.

Brain and whole body localization and distribution of 125I-leptin was determined after intraperitoneal administration to ob/ob and db/db mice, and was compared to inhibition of food intake. Food intake was not significantly inhibited at3 hours post-injection, but was decreased significantly at 6 h (p0.0007) and 24 h (p0.02) in ob/ob mice, times at which97 % of the radioactive dose was found in the urine. The highest concentrations of 125I-leptin at all time-points were found in the serum, liver and kidneys. These findings were verified by whole body autoradiography. Virtually no 125I-leptin was found in the CNS at later timepoints in either ob/ob or db/db mice. Coronal sectioning of entire brains from ob/ob and db/db mice revealed 125I radioactivity localized to the choroid plexus and in the ventricular space, but not in other CNS regions. No differences in localization, accumulation, or clearance of 125I-leptin in ob/ob vs. db/db mice were found in any of the tissues studied. The present studies demonstrate that the inhibitory effect of leptin on food intake in the ob/ob mouse persists for up to 24 hours after a single dose, despite the complete degradation and elimination of the labeled leptin during the first several hours after injection.

The structural abnormalities and functional characteristics of dysfunctional prothrombin variants in two new kindreds have been determined. Prothrombin Corpus Christi (family 1) was purified and found to have markedly reduced fibrinogen clotting activity, yet normal amidolytic and near-normal platelet aggregating activity. A transition (C to T) at nucleotide position 8885, present in the heterozygous form in affected family members, resulted in the substitution of Cys for Arg 382. This substitution results in the loss of a positive charge within the fibrinogen-binding exosite of thrombin, thus accounting for the observed functional defect. A heterozygous C to T transition was also present at position 19994 in other family members with a hypoprothrombinemic phenotype. This mutation results in the replacement of Gln 541 (CAA) by a premature stop codon (TAA). Prothrombin Dhahran (family 2) was found to have markedly reduced fibrinogen clotting activity, but normal amidolytic activity. Affected family members were found to have a G to A transition at nucleotide position 7312 resulting in the substitution of His for Arg 271. This substitution results in the abolition of a factor Xa cleavage site, yielding meizothrombin rather than thrombin, on activation of prothrombin Dhahran by factor Xa. All but one of the above mutations occur at CpG dinucleotides, thus further supporting the observation of a high incidence of CpG transitions in hereditary dysprothrombinemia. The significant bleeding tendencies of individuals homozygous for prothrombin Dhahran (prothrombin clotting activity 5% to 7%) contrast sharply with the absence of significant chronic bleeding in the proband expressing prothrombin Corpus Christi (prothrombin clotting activity 2%). Our findings underscore the capacity of thrombin to contribute to clinical hemostasis by mechanisms other than its fibrinogen clotting activity.

Fibro-osseous pseudotumor of the digit is an unusual cutaneous process characterized histologically by a fibroblastic proliferation admixed with reactive/metaplastic osteoid formation. The osteoid formation can be florid and immature, mimicking the appearance of malignant osteoid-forming neoplasms. Fibro-osseous pseudotumor of the digit has histologic and clinical features in common with myositis ossificans. This has led many to consider the two to be synonymous. We studied three cases of fibro-osseous pseudotumor, compared to five cases of myositis ossificans, using routine light microscopy and a battery of immunohistochemical stains. Both entities displayed a "zoning" pattern of immature spindled areas admixed with more mature areas having osteoid metaplasia. This was more pronounced in myositis ossificans. In each lesion, the spindle cells stained positively for vimentin and actin. CD34 and Factor VIII highlighted the vasculature. No stromal staining for MAK-6 (cytokeratin) or S-100 was identified. Ki-67, a proliferation marker, showed positive staining of the stromal cells in both lesions, which was strongest in the immature spindled areas. The immunohistochemical and histologic similarities of the lesions support fibro-osseous pseudotumor of the digit being a cutaneous variant of myositis ossificans.

A CryV-type protein (CGCryV) has been isolated from supernatant fluids of Bacillus thuringiensis AB88 cultures. Previous reports have suggested the cryptic nature of the cryV-type genes on the basis of the absence of CryV-type proteins in parasporal crystals. The CryV-type protein reported here is expressed early in stationary phase, and evidence indicates that it is an exported protein. Analysis of the deduced protein sequence from this gene reveals the presence of an N-terminal domain that likely acts as a signal peptide. The CGCryV protein is the first reported case of a delta-endotoxin being a secreted protein, which may influence the biological relevance of these proteins.

Nitric oxide is a significant biologic mediator in a number of physiologic processes. Clinical and laboratory studies in both human and animal models have uncovered a number of conditions responsive to nitric oxide therapy. The use of inhaled nitric oxide is rapidly expanding into neonatal and pediatric critical care. Presently, the primary clinical indication for nitric oxide is pulmonary hypertension of either a primary or secondary etiology. Some patient populations that are refractory to conventional management demonstrate significant improvement when receiving nitric oxide therapy. This article discusses the physiologic properties of nitric oxide, as well as its diagnostic and therapeutic indications. Specific issues regarding nitric oxide delivery, monitoring, safety standards, and nursing care are also addressed.

The Sabie River is an important river catchment in South Africa because of its ecodiversity, importance to the Kruger National Park, agricultural and afforestation potential and relatively undeveloped water resources. These resources are under stress from increasing irrigation and a burgeoning population. In the mid-eighties an intensive river basin study was undertaken to guide and promote sustained development. The studies described in this paper were undertaken in order to include economic criteria in development investment decision making. These studies were firstly, a macro-economic view which extended to sector analysis and secondly, a cost-benefit analysis of the proposals. The first found that the balance between economic growth and the demand for water takes place in an area which exhibits a dualistic and interdependent nature, between the Sabie River subregion with dynamic economic growth sectors, and the Sand River subregion with a small economic base but high population concentration. The study introduces the concept of investment absorption capacity which uses Gross Geographic Product as a proxy in a methodology that indicates the maximum capital that can, technically, be invested in water resource development in the area. The second part uses traditional cost-benefit analysis to determine the economically preferred proposals for development.

The synthesis and spectroscopic properties of all the mono-, di-, tri-, tetra-, penta-, hexa-, and heptachlorobipheynls are reported and the synthesis of all 209 polychlorinated biphenyls (PCBs) is completed. The retention times and molar response factors of the 209 PCBs were determined relative to a reference standard, octachloronaphthalene. The retention times for these compounds generally increased with increasing chlorine content, and it was apparent that within a series of isomers there was an increase in retention time with increasing meta and para and decreasing ortho substitution. By use of a 50-m narrow bore fused silica capillary column coated with SE-54, it was possible to separate 187 PCB congeners, and only 11 pairs of compounds were not fully resolved. With some additional analytical improvements, isomer-specific PCB analysis can be utilized to determine the composition of commercial PCBs and accurately follow the fate and distribution of these pollutants within the global ecosystem.

Multiple myeloma (MM) is an incurable, B-cell malignancy characterized by the clonal proliferation and accumulation of malignant plasma cells in bone marrow. Despite recent advances in the understanding of genomic aberrations, a comprehensive catalogue of clinically actionable mutations in MM is just beginning to emerge. The tyrosine kinase (TK) and RAS oncogenes, which encode important regulators of various signaling pathways, are among the most frequently altered gene families in cancer. To clarify the role of TK and RAS genes in the pathogenesis of MM, we performed a systematic, targeted screening of mutations on prioritized RAS and TK genes, in CD138-sorted bone marrow specimens from 42 untreated patients. We identified a total of 24 mutations in the KRAS, PIK3CA, INSR, LTK, and MERTK genes. In particular, seven novel mutations in addition to known KRAS mutations were observed. Prediction analysis tools PolyPhen and Sorting Intolerant from Tolerant (SIFT) were used to assess the functional significance of these novel mutations. Our analysis predicted that these mutations may have a deleterious effect, resulting in the functional alteration of proteins involved in the pathogenesis of myeloma. While further investigation is needed to determine the functional consequences of these proteins, mutational testing of the RAS and TK genes in larger myeloma cohorts might also be useful to establish the recurrent nature of these mutations.

Conservation and management of natural populations requires accurate and inexpensive genotyping methods. Traditional microsatellite, or simple sequence repeat (SSR), marker analysis remains a popular genotyping method because of the comparatively low cost of marker development, ease of analysis and high power of genotype discrimination. With the availability of massively parallel sequencing (MPS), it is now possible to sequence microsatellite-enriched genomic libraries in multiplex pools. To test this approach, we prepared seven microsatellite-enriched, barcoded genomic libraries from diverse taxa (two conifer trees, five birds) and sequenced these on one lane of the Illumina Genome Analyzer using paired-end 80-bp reads. In this experiment, we screened 6.1 million sequences and identified 356 958 unique microreads that contained di- or trinucleotide microsatellites. Examination of four species shows that our conversion rate from raw sequences to polymorphic markers compares favourably to Sanger- and 454-based methods. The advantage of multiplexed MPS is that the staggering capacity of modern microread sequencing is spread across many libraries; this reduces sample preparation and sequencing costs to less than $400 (USD) per species. This price is sufficiently low that microsatellite libraries could be prepared and sequenced for all 1373 organisms listed as ‘threatened’ and ‘endangered’ in the United States for under $0.5 M (USD).

Individuals with autism spectrum disorders (ASD) often exhibit impaired executive function (EF) performance, including difficulty with working memory (WM), in particular. While research has documented the existence of these deficits, surprisingly little research exists that evaluates potential treatment strategies for improving EF or WM. One exception is a study that used positive reinforcement to improve performance on a classical WM task, the counting span, resulting in both maintenance and generalization ( Baltruschat et al., 2011 ). The current study is the second in a programmatic line of research on behavioral intervention for improving WM in children with autism. This study extended the use of the same procedure (positive reinforcement) to another task which is said to measure WM, a Complex Span, and included three additional children with autism. Results demonstrated significant improvements in performance for each participant, including maintenance and generalization to untrained stimuli and untrained responses. These results provide further evidence that behavioral intervention procedures may be useful for improving skills labeled as EF or WM in children with ASD.