STR multiplexes have been indispensable for the efficient genotyping of forensic samples. The PowerPlex 16 System contains the coreCODIS loci, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, ...D18S51, D21S11, CSF1PO, FGA, THOI, TPOX, vWA, the sex determinant locus, amelogenin, and two pentanucleotide STR loci, Penta D and Penta E. This multiplex satisfies the locus requirements for most national databases and is the most efficient currently available system due to its single PCR amplification. To provide the groundwork for judicial acceptance, including the publication of primer sequences, and to evaluate laboratory-to-laboratory variation, a developmental validation for casework on this commercially available system was performed in 24 laboratories and produced the following conclusions. Amplification was reliable on a variety of thermal cyclers and product could be analyzed on either an ABI PRISM 310 Genetic Analyzer or an ABI PRISM 377 DNA Sequencer. Genotyping using single source samples was consistent between 0.25 and 2 ng of input DNA template with a few laboratories obtaining complete genotypes at 0.0625 ng. However, heterozygote allele imbalance (<60% peak height balance) caused by stochastic effects was observed at a rate of 13% with 0.125 ng DNA and 22% at 0.0625 ng DNA. Mixture analyses were done using a total of 1 ng of DNA template. Most alleles were detected in mixtures of 4 to 1 and some minor alleles were detected in mixtures of 19 to 1. Optimum amplification cycle number was dependent on the sensitivity of the detection instrument used and could also be adjusted to accommodate larger amounts of DNA on solid supports such as FTA paper. Reaction conditions including volume, annealing temperature, and concentrations of primer, AmpliTaq Gold, and magnesium were shown to be optimal yet robust enough to withstand moderate variations without affecting genotype analysis. Environmental, matrix and standard source analyses revealed an ability to obtain complete genotypes in all sample types except those exposed to 80 degrees C for 12-48 days. Finally, comparison of genotype results from the PowerPlex 16 System with other commercially available systems on non-probative reference and forensic samples showed consistent results.
Sarracenia purpurea L., a carnivorous bog plant (also known as the pitcher plant), represents an excellent model of a well-defined, self-contained ecosystem; the individual pitchers of the plant ...serve as a microhabitat for a variety of micro- and macro-organisms. Previously, fecal indicator bacteria (Escherichia coli and enterococci) were shown as incidental contaminants in pitcher fluid; however, whether their occurrence in pitcher fluid is incidental or common has not been established. The purpose of this study was to investigate the occurrence, distribution, and growth potential of E. coli and enterococci in pitcher plant fluid from a protected bog in northwest Indiana. Escherichia coli and enterococci were recovered in pitcher fluids (n = 43 plants), with mean densities (log CFU mL
-1
) of 1.28 ± 0.23 and 1.97 ± 0.27, respectively. In vitro experiments showed that E. coli growth in fluid not containing insects or indigenous organisms was directly proportional to the fluid concentration (growth was 10-fold in 24 h in 100% fluid); however, in the presence of other indigenous organisms, E. coli and enterococci were only sustained for 5 days at 26 °C. Pulsed-field gel electrophoresis (PFGE) analysis showed that the plant Enterococcus faecalis isolates were genetically distinct from the human isolates; identical PFGE patterns were observed among plant isolates that fell into one of six clonal groups. These findings suggest that (i) E. coli and enterococci occurrence in pitcher plants is rather common in the bog studied, although their originating source is unclear, and (ii) the pitcher fluid contains adequate nutrients, especially carbon and energy sources, to promote the growth of indicator bacteria; however, under natural conditions, the biotic factors (e.g., competition for nutrients) may restrict their growth.Key words: indicator bacteria, environmental occurrence, microcosm, pitcher plant fluid, temperate bog.
Background Pruritus is the hallmark clinical sign of atopic dermatitis ( AD ) in dogs. Preliminary study results suggest that oclacitinib, a selective J anus kinase inhibitor, could reduce pruritus ...and associated inflammatory skin lesions in dogs with AD . Hypothesis/Objectives The objective was to evaluate efficacy and safety of oclacitinib ( A poquel ® ) for the control of AD in a randomized, double‐blind, placebo‐controlled trial. Animals Clinicians at 18 specialty clinics enrolled client‐owned dogs ( n = 299) with a history of chronic AD . Methods Dogs were randomized to receive either oclacitinib (0.4–0.6 mg/kg twice daily for 14 days and then once daily for up to 112 days) or an excipient‐matched placebo. Owners assessed visual analog scale (VAS) scores of pruritus on days 0, 1, 2, 7, 14, 28, 56, 84 and 112. Clinicians assessed Canine AD Extent and Severity Index ( CADESI ‐02) scores on days 0, 14, 28, 56, 84 and 112. Results On days 1, 2, 7, 14 and 28, oclacitinib‐treated dogs had a 29.5, 42.3, 61.5, 66.7 and 47.4% reduction from baseline in owner‐assessed pruritus scores, respectively, compared with a 6.5, 9.1, 6.5, 3.9 and 10.4% reduction in placebo‐treated dogs. On days 14 and 28, dermatologists recorded a 48.4% reduction in CADESI ‐02 scores in oclacitinib‐treated dogs compared with a 1.7% reduction and a 3.6% increase in placebo‐treated dogs. After day 28, >86% of all placebo‐treated dogs had moved to an open‐label study, making between‐group comparisons biased. Differences were significant at all time points assessed ( P < 0.0001). Conclusions and clinical importance Oclacitinib provided rapid, effective and safe control of AD , with substantial improvement in VAS and CADESI ‐02 scores.
Resumen Introducción el prurito es la signo clínico mas relevante de la dermatitis atópica ( AD ). Estudios preliminares sugieren que oclacinitib, un inhibidor selectivo de la quinasa Janus, puede reducir el prurito y las lesiones cutáneas inflamatorias asociadas a perros con AD . Hipótesis/Objetivos el objetivo fue evaluar la eficacia y seguridad de oclacinitib ( A poquel ® ) para el control de la dermatitis atópica en un a prueba doble ciega al azar controlada con placebo. Animales veterinarios clínicos de 18 hospitales especializados reclutaron perros de propietarios particulares (n = 299) con historia de AD crónica. Métodos los perros fueron distribuidos al azar para recibir bien oclacitinib (0,4‐ 0,6 mg/kg dos veces al día durante 14 días y después una vez al día hasta por 112 días) o un placebo con el mismo excipiente. Los propiertarios valoraron en la escala análoga visual el índice de prurito en los días 0, 1, 2, 7, 14, 28, 56, 84 y 112. Los clínicos hicieron lo mismo siguiendo el índice canino de extensión y severidad del prurito ( CADESI ‐02) en los días 0, 14, 28, 56, 84 y 112. Resultados en los días 1, 2, 7, 14 y 28 los perros tratados con oclacinitib tuvieron una reducción del 29,5; 42,3; 61,5; 66,7 y 47,4% con respecto a los valores iniciales en la escala de prurito evaluada por los propietarios, respectivamente. Esto estuvo en contraste con la reducción en los perros tratados con placebo que fue de 6,5; 9,1; 6,5; 3,9 y 10,4%, respectivamente. En los días 14 y 28 los dermatólogos notaron una reducción del 48,4% en los valores de CADESI ‐02 en los perros tratados con oclacitinib comparado con una reducción de 1,7% y un aumento de 3,6% en perros tratados con placebo. Después del día 28, >86% de los perros tratados con placebo se pasaron a un estudio abierto, desviando la comparación entre grupos. Las diferencias fueron significativas en todos los puntos estudiados ( P = 0,0001). Conclusiones e importancia clínica oclacitinib produjo un control rápido, efectivo y seguro de la AD con mejora sustancial en la escala visual análoga y los valores CADESI ‐02.
Résumé Contexte Le prurit est le signe clinique typique de la dermatite atopique ( AD ) chez le chien. Les résultats d’études préliminaires suggèrent que l'oclacitinib, un inhibiteur sélectif de Janus kinase, pourrait réduire le prurit et les lésions inflammatoires cutanées associées chez les chiens atteints d’ AD . Hypothèses/Objectifs L'objectif était d’évaluer l'efficacité et l'innocuité de l'oclacitinib ( A poquel®) pour le contrôle de l’ AD dans une étude randomisée, contrôlée contre placebo, en double aveugle. Sujets Les cliniciens de 18 cliniques spécialisées ont inclus des chiens de propriétaires ( n = 299) atteints de dermatite atopique chronique. Méthodes Les chiens ont été randomisés pour recevoir soit de l'oclacitinib (0.4–0.6 mg/kg deux fois par jour pendant 14 jours puis une fois par jour pendant 112 jours) soit un placebo contenant l'excipient. Les propriétaires ont réalisé des échelles de prurit visuelles à jours 0, 1, 2, 7, 14, 28, 56, 84 et 112.Les cliniciens ont réalisé des scores CADESI ‐02 (Canine AD Extent and Severity Index) à jours 0, 14, 28, 56, 84 et 112. Résultats Aux jours 1, 2, 7, 14 et 28, les chiens ayant reçu de l'oclacitinib avaient respectivement 29.5, 42.3, 61.5, 66.7 et 47.4% de réduction de scores de prurit évalué par les propriétaires comparé à 6.5, 9.1, 6.5, 3.9 et 10.4% de diminution chez les chiens contrôles ayant reçu le placebo. A jours 14 et 28, les dermatologues ont enregistré une diminution de 48.4% des scores CADESI ‐02 chez les chiens recevant de l'oclacitinib comparé à une diminution de 1.7% et une augmentation de 3.6% chez les chiens recevant le placebo. Après le jour 28, plus de 86% des chiens recevant le placebo ont été transféré dans une étude ouverte faisant des biais de comparaison entre les groupes. Les différences étaient significatives à tous les points évalués ( P < 0.0001). Conclusions et importance clinique L'oclacitinib a entrainé un contrôle rapide, efficace et sûr de l’ AD , avec une amélioration substantielle des scores de l’échelle analogue visuelle et du CADESI ‐02.
Zusammenfassung Hintergrund Juckreiz ist bei Hunden das Kardinalsymptom der atopischen Dermatitis ( AD ). Vorläufige Studienergebnisse zeigen, dass Oclacitinib, ein selektiver Janus Kirasehemmer, den Juckreiz und damit verbundene entzündliche Hautreaktionen bei Hunden mit AD reduzieren konnte. Hypothese/Ziele Das Ziel dieser Studie war es, die Wirksamkeit und Sicherheit von Oclacitinib (Apoquel®) zur Kontrolle von AD in einer randomisierten, doppelblinden, Plazebo‐kontrollierten Studie zu evaluieren. Tiere KlinikerInnen aus 18 Spezialkliniken nahmen Hunde im Besitz von KundInnen (n = 299) mit der Anamnese einer chronischen AD in die Studie auf. Methoden Die Hunde erhielten zufällig entweder Oclacitinib (0,4‐0,6 mg/kg zweimal täglich für 14 Tage und dann einmal täglich für bis zu 112 Tage) oder ein dem Trägermedium angepasstes Plazebo. Die BesitzerInnen beurteilten die Juckreiz Werte mittels visueller Analogskala an den Tagen 0, 1, 2, 7, 14, 28, 84 und 112. Die KlinikerInnen beurteilten die Canine AD Extent and Severity Index Werte ( CADESI ‐02) an den Tagen 0, 14, 28, 56, 84 und 112. Ergebnisse An den Tagen 1, 2, 7, 14 und 28 zeigten die mit Oclacitinib behandelten Hunde eine 29,5; 42,3; 61,5 bzw 47,4%ige Reduktion der von den BesitzerInnen beurteilten Juckreizwerte im Vergleich zu 6,5; 9,1; 6,5; 3,9 und 10,4%igen Reduktion bei den mit Plazebo behandelten Hunden. An den Tagen 14 und 28 beschrieben die DermatologInnen eine 48,4%ige Reduktion der CADESI‐02 Werte bei den mit Oclacitinib behandelten Hunde im Vergleich zu einer 1,7%igen Reduktion und einem 3,6%igen Anstieg bei den mit Plazebo behandelten Hunde. Nach dem Tag 28 waren >86% aller Plazebo‐behandelten Hunde in einer offenen Studie, was die Vergleiche zwischen den Gruppen polarisiert. Die Unterschiede waren zu allen Zeitpunkten signifikant verschieden (P<0,0001). Schlussfolgerungen und klinische Bedeutung Oclacitinib stellte eine rasche, wirksame und sichere Kontrolle von AD mit einer deutlichen Verbesserung der Werte der visuellen Analogskala und der CADESI ‐02 Werte dar.
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Eighteen adult subjects with mild to moderate hepatic impairment and 18 healthy control subjects were given a single 15‐mg dose of sirolimus by oral solution. Mean whole‐blood sirolimus ...weight‐normalized oral‐dose clearances (CL/F) were significantly decreased (P = .02) in subjects with mild to moderate hepatic impairment by −31.8% and −36.0%, respectively, compared with controls. There were no significant differences in mean sirolimus Cmax and tmax values among groups. The observed decreases in CL/F may be relevant in renal transplant patients with mild to moderate hepatic impairment, based on the close similarity of sirolimus CL/F in controls and previously studied stable renal transplant patients receiving multiple‐dose administration of sirolimus and cyclosporine. There was considerable overlap in the CL/F values of hepatic‐impaired subjects and controls, suggesting that whole‐blood sirolimus trough concentrations in renal transplant patients exhibiting mild to moderate hepatic impairment be initially monitored to assess the need for dose adjustments.
The mobile group II intron of
Lactococcus lactis, Ll.LtrB, provides the opportunity to analyze the homing pathway in genetically tractable bacterial systems. Here, we show that Ll.LtrB mobility ...occurs by an RNA-based retrohoming mechanism in both
Escherichia coli and
L. lactis. Surprisingly, retrohoming occurs efficiently in the absence of RecA function, with a relaxed requirement for flanking exon homology and without coconversion of exon markers. These results lead to a model for bacterial retrohoming in which the intron integrates into recipient DNA by complete reverse splicing and serves as the template for cDNA synthesis. The retrohoming reaction is completed in unprecedented fashion by a DNA repair event that is independent of homologous recombination between the alleles. Thus, Ll.LtrB has many features of retrotransposons, with practical and evolutionary implications.
Abstract only
The spread of insect‐transmitted diseases is prevalent due to the female blood‐feeding mosquitos’ ability to carry pathogens. The iron component of the blood meal is critical for ...mosquito oogenesis, and thus, the manipulation of iron metabolism is a potential option to develop mosquito control strategies and decrease the transmission of these diseases. A major protein in iron metabolism is iron regulatory protein 1 (IRP1). IRP1 controls translation of several proteins involved in iron metabolism. In mammals, phosphorylation occurs at two sites by protein kinase C (PKC) and alters IRP1 interaction with mRNA. Mosquito IRP1 contains one site homologous to the mammalian IRP1 as well as a second unique site. Phosphorylation at the unique site may explain the different behavior of mosquito IRP1. This study attempts to determine if mosquito IRP1 is phosphorylated.
Aedes aegypti (
yellow fever mosquito) CCL‐125 cells or
Anopheles gambiae
4a3b cells were treated with phorbol‐12‐myristate‐13‐acetate (PMA) to stimulate PKC. We report our efforts to detect IRP1 phosphorylation in these cells using SDS‐PAGE, fluorescent staining and treatment of the cells in the presence of
32
P‐orthophosphate followed by immuno‐precipitation. Experiments using mammalian cells served as controls. In the long term, this investigation will contribute to the solution of mosquito population control and reduction of infectious disease.
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