Development of a Screening Algorithm for Lung Disease in Systemic Juvenile Idiopathic Arthritis.

OBJECTIVE: Lung disease (LD) is an increasingly recognized complication of systemic juvenile idiopathic arthritis (sJIA). As there are no currently available guidelines for pulmonary screening in sJIA, we sought to develop such an algorithm at our institution. METHODS: A multidisciplinary workgroup was convened, including members representing rheumatology, pulmonary, stem cell transplantation, and patient families. The workgroup leaders drafted an initial algorithm based on published literature and experience at our center. A modified Delphi approach was used to achieve agreement through three rounds of anonymous, asynchronous voting and a consensus meeting. Statements approved by the workgroup were rated as appropriate with moderate or high levels of consensus. These statements were organized into the final approved screening algorithm for LD in sJIA. RESULTS: The workgroup ultimately rated 20 statements as appropriate with a moderate or high level of consensus. The approved algorithm recommends pulmonary screening for newly diagnosed patients with sJIA with clinical features that the workgroup agreed may confer increased risk for LD. These "red flag features" include baseline characteristics (young age of sJIA onset, human leukocyte antigen type, trisomy 21), high disease activity (macrophage activation syndrome [MAS], sJIA-related ICU admission, elevated MAS biomarkers), respiratory symptoms or abnormal pulmonary examination findings, and features of drug hypersensitivity-like reactions (eosinophilia, atypical rash, anaphylaxis). The workgroup achieved consensus on the recommended pulmonary work-up and monitoring guidelines. CONCLUSION: We developed a pulmonary screening algorithm for sJIA-LD through a multidisciplinary consensus-building process, which will be revised as our understanding of sJIA-LD continues to evolve.

Common Whitethroats Curruca communis show a continuum of residency duration but a high degree of between-years site fidelity at nonbreeding grounds in Nigeria.

The nonbreeding period represents a significant part of an Afro-Palearctic migratory bird's annual cycle. Decisions such as whether to remain at a single site and whether to return to it across years have important effects on aspects such as survival, future breeding success, migratory connectivity, and conservation. During this study, we color-ringed 337 common Whitethroats Curruca communis and undertook daily resightings to understand site persistence and the degree of site fidelity throughout three nonbreeding periods (November-April) in Nigeria. The probability of detecting a color-ringed Whitethroat when it was present was 0.33. Site persistence varied widely across individuals (1-165 days) and did not differ significantly with sex or year, though first-year birds remained for significantly shorter periods than adults. We believe that shorter residencies are likely due to the use of multiple stationary nonbreeding sites rather than low winter survival. A minimum of 19% of individuals returned to the study site the following year and shifted, on average, 300 m, suggesting that Whitethroats have a relatively high degree of between-years site fidelity at a very fine scale. An individual's previous residency duration did not seem to determine its residency duration the following year. We suggest that spatial fidelity is high and constant through years, but temporal fidelity is not, and individual residency patterns vary, probably according to yearly and seasonal conditions. Our results highlight the complexity of the annual cycle of a single species and the importance of carrying out in situ, fine-scale research throughout a migrant's annual cycle over several years.

Migratory routes, breeding locations and multiple non-breeding sites of Common Whitethroats Curruca communis revealed by geolocators.

Understanding general migration characteristics and how breeding and non-breeding sites are connected is crucial for predicting the response of long-distance migratory bird populations to environmental changes. We use data collected from six geolocators to describe migratory routes and identify breeding and non-breeding locations, migratory behaviour and differences between spring and autumn migration of Common Whitethroats Curruca communis, an Afro-Palearctic migrant, wintering in Nigeria. Most individuals departed on spring migration in April, following a north-easterly direction, arriving at their breeding grounds across central-eastern Europe (~425,000 km2) in May. Departures from breeding grounds took place between July and August in a south-westerly direction. During spring migration individuals travelled longer distances at faster rates making its overall duration shorter than autumn migration. We suggest that, while Whitethroats can cross the Sahara Desert and Mediterranean Sea in a single flight, they are likely to refuel before and after crossing. Results indicate that Whitethroats undertook loop migration and visited two wintering sites: first in the Sahel, then in Nigeria, where they remained until spring migration. Geolocator results and data from the European Union for Bird Migration's (EURING) ringing database suggest that Whitethroats have a relatively high migratory spread-individuals from a single non-breeding site breed across a wide area of Europe. Our research is the first to track and describe the complete annual cycle of Whitethroats and one of the few studies to do so for any Afro-Palearctic migrant from non-breeding grounds. We identified the Sahel as an important refuelling and first wintering site indicating its conservation, alongside other stopover sites, is crucial for the species. We believe that changes in this region will have severe effects on a subset of individuals of specific European breeding populations, but these effects will greatly depend on the severity of the changes and at what spatial scale they occur.

Rapid quantification of the aminoglycoside arbekacin in serum using high performance liquid chromatography-tandem mass spectrometry.

BACKGROUND: This project entails the development and validation of a method for quantification of the aminoglycoside antibiotic arbekacin in serum using liquid chromatography tandem mass spectrometry (LC-MS/MS) for therapeutic drug monitoring in future clinical trials. METHODS: Following a protein precipitation with 0.3 mol/l perchloric acid containing internal standard dibekacin at a concentration of 0.6 µg/ml, human serum samples containing arbekacin were analyzed using a Hypersil Gold PFP column and a liquid chromatography system. Elution occurred with a gradient of water and acetonitrile, each containing 0.05% (v/v) trifluoroacetic acid and 0.1% (v/v) formic acid. Analytes were detected over a 3.25 minute run time using a tandem mass spectrometer with a heated electrospray-ionization (HESI) source in positive ionization mode with selected reaction monitoring (SRM). Matrix effects, carryover, linearity, recovery, precision, and limit of quantification were carefully evaluated. RESULTS: The limit of quantification for arbekacin was 0.1 µg/ml. All simple and total precision CV's were less than 6.2%. The method was linear from 0.1 µg/ml to 45.9 µg/ml (slope of 0.973). The mean recovery ranged from 94.7 to 103.8%. No matrix effects were detected. CONCLUSIONS: This developed and validated LC-MS/MS method allows for the quantification of arbekacin in serum following protein precipitation.

Determining clinical and psychological benefits and barriers with continuous glucose monitoring therapy.

BACKGROUND: Preventing the complications of diabetes requires tight control and minimizing blood glucose fluctuation. Pursuing these goals increases the risk of severe hypoglycemia. The authors hoped to identify if using continuous glucose monitoring (CGM) decreased the incidence of severe hypoglycemia, resulted in less fear of hypoglycemia, and improved patient empowerment and what impact CCM had on quality of life (QOL) issues. METHODS: Questionnaires were used to gather demographic data, to measure educational experiences with CGM, QOL issues concerning fear of hypoglycemia and severe hypoglycemia, confidence to make changes to insulin regimens before and after using CGM, and the incidence of severe hypoglycemia before and while using CGM, and to assess the impact of CGM on QOL. RESULTS: The data and answers reported suggest significantly less fear of hypoglycemia and severe hypoglycemia and increased patient empowerment with CGM. Further shown is a reduced incidence of acute complications as evidenced by significantly decreased incidence of severe hypoglycemia. CGM appears to have a significant, positive impact on the stress associated with having and managing diabetes. CONCLUSIONS: Using CGM as part of the diabetes self-management plan offers the potential to significantly improve user outcomes as measured by QOL, reduction of fear, and patient empowerment. Using CGM can allow patients to achieve tighter control of their blood glucose values with reduced fear of hypoglycemia, reduced incidence of severe hypoglycemia, and a decreased sense of burden from having diabetes.

Addition of epidermal growth factor improves the rate of sulfur mustard wound healing in an in vitro model.

OBJECTIVE: Sulfur mustard (SM) causes blisters on the human skin. These blisters delay healing of the skin and make the victims more susceptible to infection. In vitro models have been used for protection studies against SM injury, but study on wound healing after SM exposure has not been explored. The purpose of this study was to test whether the addition of exogenous growth factors could improve the rate of SM wound healing. METHODS: The model consisted of normal human epidermal keratinocytes seeded into 6-well plates, exposed to SM, and wounded (disruption of the cell monolayer) with a sterile wounding instrument. Cells were then stained and images were captured to measure percentage wound fill. Epidermal growth factor (EGF) and keratinocyte growth factor (KGF) were tested in this model. RESULTS: EGF (1 ng/mL) significantly increased wound fill on all of the days tested (days 6, 9, and 12). KGF did not significantly improve wound healing. CONCLUSIONS: EGF showed promise as a potential therapy for SM-induced wounds. This in vitro model was a valuable tool for screening therapeutics before animal testing. These results will be used to develop a dressing that can slowly release EGF on to a debrided wound bed to help speed the healing process.

Sex differences in the influence of COMT Val158Met on alcoholism and smoking in plains American Indians.

BACKGROUND: Alcoholism and heavy smoking are highly comorbid and are cotransmitted in the general U.S. population; however little is known about comorbidity in American Indians. The catechol-O-methyltransferase (COMT) functional polymorphism, Val158Met, has been associated with alcoholism in Caucasians. The aims of our study were firstly to investigate patterns of alcohol and tobacco consumption and comorbidity between alcoholism and smoking in Plains American Indians and secondly to determine the influence, including sexual dimorphic effects, of COMT Val158Met and COMT haplotypes, on these behaviors. METHODS: Diagnostic and Statistical Manual-III-R lifetime diagnoses were assigned to 342 community-ascertained Plains American Indians (201 women, 141 men). Lifetime drinking and smoking histories were obtained. Five COMT loci, including Val158Met, were genotyped. Haplotype-based analyses identified 1 block with 3 common haplotypes; 2 included Val158, and 1 had the Met158 allele. RESULTS: The alcoholics drank heavily (12+/-8 drinks/drinking day) but episodically (max 10+/-8 d/mo). Although 62% of male alcoholics and 40% of female alcoholics were smokers (> or =10 cigarettes/d), only 12% of alcoholic men and 8% of alcoholic women smoked heavily (>20/d). In women, the COMT Val158 allele frequency was maximal in alcoholic smokers (0.85), decreasing to 0.74 in nonalcoholic smokers, 0.67 in alcoholic nonsmokers, and 0.64 in nonalcoholic nonsmokers (chi2 = 11.1, 3 df, p = 0.011). Women showed a main effect of Val158 on smoking (p=0.003). Both male and female alcoholics were more likely to have at least 1 Val158 allele compared with nonalcoholics (0.95 vs 0.88, p < 0.05). Approximately 30% of all participants were long-term, nonaddicted light, social smokers (3.6+/-1.7 cigarettes/d); they had the same Val158Met frequencies as nonsmokers. Haplotype analyses supported the Val158Met findings; however, only 1 of the 2 Val158 haplotypes was implicated. CONCLUSIONS: Plains Indians have different smoking and drinking patterns and considerably less comorbidity between alcoholism and heavy smoking compared with the general U.S. population. Our COMT Val158Met results suggest that there may be both sex differences in the genetic origins of alcoholism and smoking in this population and overlap in genetic vulnerability to both addictions in women.

Microarray analysis of mouse ear tissue exposed to bis-(2-chloroethyl) sulfide: gene expression profiles correlate with treatment efficacy and an established clinical endpoint.

Bis-(2-chloroethyl) sulfide (sulfur mustard; SM) is a potent alkylating agent. Three treatment compounds have been shown to limit SM damage in the mouse ear vesicant model: dimercaprol, octyl homovanillamide, and indomethacin. Microarrays were used to determine gene expression profiles of biopsies taken from mouse ears after exposure to SM in the presence or absence of treatment compounds. Mouse ears were topically exposed to SM alone or were pretreated for 15 min with a treatment compound and then exposed to SM. Ear tissue was harvested 24 h after exposure for ear weight determination, the endpoint used to evaluate treatment compound efficacy. RNA extracted from the tissues was used to generate microarray probes for gene expression profiling of therapeutic responses. Principal component analysis of the gene expression data revealed partitioning of the samples based on treatment compound and SM exposure. Patterns of gene responses to the treatment compounds were indicative of exposure condition and were phenotypically anchored to ear weight. Pretreatment with indomethacin, the least effective treatment compound, produced ear weights close to those treated with SM alone. Ear weights from animals pretreated with dimercaprol or octyl homovanillamide were more closely associated with exposure to vehicle alone. Correlation coefficients between gene expression level and ear weight revealed genes involved in mediating responses to both SM exposure and treatment compounds. These data provide a basis for elucidating the mechanisms of response to SM and drug treatment and also provide a basis for developing strategies to accelerate development of effective SM medical countermeasures.

BDNF variation and mood disorders: a novel functional promoter polymorphism and Val66Met are associated with anxiety but have opposing effects.

The brain-derived neurotrophic factor (BDNF) gene is critical for neuronal function and survival, and is likely to be important in psychiatric disorders. In this study, we used single-nucleotide polymorphism (SNP) discovery, functional analyses, and genetic association studies to better understand the potential role of BDNF sequence variation in behavior. Screening 480 unrelated individuals for SNPs and genotyping was performed in US Caucasian, American Indian, and African American populations. Lifetime DSM-III-R psychiatric diagnoses were assigned and the Tridimensional Personality Questionnaire (TPQ) was administered to measure anxious temperament (harm avoidance (HA)) and novelty seeking (NS). A novel SNP (-281 C>A) in promoter 1 was discovered that had decreased DNA binding in vitro and decreased basal reporter gene activity in transfected rat hippocampal neurons. The frequency of the -281 A allele was 0.03 in a Caucasian sample, but was virtually absent in other populations. Association analyses in a community-based sample showed that individuals with the -281 A allele (13 heterozygotes) had lower TPQ HA (F=4.8, p<0.05). In contrast, the Met 66 allele was associated with increased HA (F=4.1, p=0.02) and was most abundant in individuals with both anxiety disorders and major depression (p<0.05). Among the Val66Val homozygotes, individuals who were -281 CA heterozygotes had significantly lower HA than the -281 CC homozygotes (p<0.01). Our results suggest that in this population, the low activity -281 A allele may be protective against anxiety and psychiatric morbidity, whereas Met 66 may be a risk allele.

Failure to detect DUP25 in lymphoblastoid cells derived from patients with panic disorder and control individuals representing European and American populations.

Investigation of the co-occurrence of panic and phobic disorders with joint laxity led to the identification of interstitial duplications involving human chromosome 15q24-26 (named 'DUP25') in a Spanish population. DUP25 was observed in 97% of patients and in 7% of control individuals. In the present study, we used two different methods to detect DUP25: high-throughput molecular gene dosage analysis and fluorescence in situ hybridization (FISH). We evaluated 56 lymphoblastoid cell lines derived from 26 unrelated patients with panic disorder obtained from several European and American populations and 30 normal controls. We could not find any cell line showing a result consistent with DUP25. These data do not support any association of DUP25 with panic disorder.

Genetic origins of anxiety in women: a role for a functional catechol-O-methyltransferase polymorphism.

OBJECTIVE: Women are more prone to anxiety than men. The catechol- -methyltransferase functional polymorphism, Val158Met, is likely to be implicated in anxiety vulnerability. We hypothesized that, particularly in women, the low-activity Met158 allele would be associated with higher anxiety scores and a biological trait, low-voltage alpha resting electroencephalogram (EEG), previously associated with alcoholism and anxiety disorders. METHODS: DNA was obtained from two independent groups of participants ascertained as community samples: 149 predominantly Caucasian individuals (92 women, 57 men), and 252 Plains American Indians (149 women, 103 men). Dimensional measures of anxiety (Tridimensional Personality Questionnaire harm avoidance subscales HA1 and HA2) were obtained and DSM-III-R lifetime psychiatric diagnoses were determined. EEGs were recorded and EEG phenotypes assigned. RESULT: In both populations, women showed significant associations between catechol- -methyltransferase genotype and elevated harm avoidance scores, and the Met158/Met158 genotype was most strongly associated: predominantly Caucasian participants: HA1, P=0.03, HA2, P =0.03; and Plains American Indians: HA2, P=0.01. This was also the case with low-voltage alpha resting EEG: predominantly Caucasian participants: P=0.01, odds ratio=5.0 (95% confidence interval, 1.3-18.7); Plains American Indians: P=0.03, odds ratio=3.7 (95% confidence interval, 1.1-12.7). CONCLUSIONS: The results of the present study suggest that an inherited difference in catecholamine metabolism is important in the pathogenesis of anxiety in women.

The relationship between two intermediate phenotypes for alcoholism: low voltage alpha EEG and low P300 ERP amplitude.

OBJECTIVE: There is considerable evidence that the amplitude of the heritable P300 event-related potential (ERP) is reduced in alcoholics and their alcohol-naive children. Low voltage alpha (LVA), a heritable resting electroencephalogram (EEG) trait present in 7-14% of the population, has been shown to be associated with alcoholism and anxiety disorders. A few studies have demonstrated a modest correlation between pre-stimulus alpha power and P300 amplitude. We aimed to test this finding in community volunteers, hypothesizing that LVA would be associated with low P300 amplitude. METHOD: Digitized resting EEG was recorded at the central parietal site (Pz) from 85 male and 113 female community volunteers (120 unrelated). ERPs were elicited at Pz by auditory and visual oddball paradigms. All participants were interviewed with the Schedule for Affective Disorders, Lifetime Version (SADS-L) and assigned blind-rated psychiatric diagnoses according to the American Psychiatric Association DSM-III-R criteria. RESULTS: LVA participants (including alcoholics and nonalcoholics) had significantly lower auditory and visual P300 amplitudes. Absolute alpha power was modestly correlated with auditory and visual P300 amplitude and was associated with 9.4% and 4.6% of the variance, respectively. CONCLUSIONS: The association between LVA and low P300 amplitude, two distinct electrophysiological traits, suggests that, at least in individuals with the LVA trait, some aspects of resting, unstimulated brain activity and activated brain function in the form of attentional response may be fundamentally related.