Synthesis and evaluation of halogenated nitrophenoxazinones as nitroreductase substrates for the detection of pathogenic bacteria

The synthesis and microbiological evaluation of 7-, 8- and 9-nitro-1,2,4-trihalogenophenoxazin-3-one substrates with potential in the detection of nitroreductase-expressing pathogenic microorganisms are described. The 7- and 9-nitrotrihalogenophenoxazinone substrates were reduced by most Gram negative microorganisms and were inhibitory to the growth of certain Gram positive bacteria; however, the majority of Gram positive strains that were not inhibited by these agents, along with the two yeast strains evaluated, did not reduce the substrates. These observations suggest there are differences in the active site structures and substrate requirements of the nitroreductase enzymes from different strains; such differences may be exploited in the future for differentiation between pathogenic microorganisms. The absence of reduction of the 8-nitrotrihalogenophenoxazinone substrates is rationalized according to their electronic properties and correlates well with previous findings

Targeted single molecule sequencing methodology for ovarian hyperstimulation syndrome.

BACKGROUND: One of the most significant issues surrounding next generation sequencing is the cost and the difficulty assembling short read lengths. Targeted capture enrichment of longer fragments using single molecule sequencing (SMS) is expected to improve both sequence assembly and base-call accuracy but, at present, there are very few examples of successful application of these technologic advances in translational research and clinical testing. We developed a targeted single molecule sequencing (T-SMS) panel for genes implicated in ovarian response to controlled ovarian hyperstimulation (COH) for infertility. RESULTS: Target enrichment was carried out using droplet-base multiplex polymerase chain reaction (PCR) technology (RainDance®) designed to yield amplicons averaging 1 kb fragment size from candidate 44 loci (99.8% unique base-pair coverage). The total targeted sequence was 3.18 Mb per sample. SMS was carried out using single molecule, real-time DNA sequencing (SMRT® Pacific Biosciences®), average raw read length = 1178 nucleotides, 5% of the amplicons \u3e6000 nucleotides). After filtering with circular consensus (CCS) reads, the mean read length was 3200 nucleotides (97% CCS accuracy). Primary data analyses, alignment and filtering utilized the Pacific Biosciences® SMRT portal. Secondary analysis was conducted using the Genome Analysis Toolkit for SNP discovery l and wANNOVAR for functional analysis of variants. Filtered functional variants 18 of 19 (94.7%) were further confirmed using conventional Sanger sequencing. CCS reads were able to accurately detect zygosity. Coverage within GC rich regions (i.e.VEGFR; 72% GC rich) was achieved by capturing long genomic DNA (gDNA) fragments and reading into regions that flank the capture regions. As proof of concept, a non-synonymous LHCGR variant captured in two severe OHSS cases, and verified by conventional sequencing. CONCLUSIONS: Combining emulsion PCR-generated 1 kb amplicons and SMRT DNA sequencing permitted greater depth of coverage for T-SMS and facilitated easier sequence assembly. To the best of our knowledge, this is the first report combining emulsion PCR and T-SMS for long reads using human DNA samples, and NGS panel designed for biomarker discovery in OHSS

Three Component Velocity and Acceleration Measurement Using FLEET

The femtosecond laser electronic excitation and tagging (FLEET) method has been used to measure three components of velocity and acceleration for the first time. A jet of pure N2 issuing into atmospheric pressure air was probed by the FLEET system. The femtosecond laser was focused down to a point to create a small measurement volume in the flow. The long-lived lifetime of this fluorescence was used to measure the location of the tagged particles at different times. Simultaneous images of the flow were taken from two orthogonal views using a mirror assembly and a single intensified CCD camera, allowing two components of velocity to be measured in each view. These different velocity components were combined to determine three orthogonal velocity components. The differences between subsequent velocity components could be used to measure the acceleration. Velocity accuracy and precision were roughly estimated to be +/-4 m/s and +/-10 m/s respectively. These errors were small compared to the approx. 100 m/s velocity of the subsonic jet studied

Optimizing the Distribution of Leg Muscles for Vertical Jumping.

A goal of biomechanics and motor control is to understand the design of the human musculoskeletal system. Here we investigated human functional morphology by making predictions about the muscle volume distribution that is optimal for a specific motor task. We examined a well-studied and relatively simple human movement, vertical jumping. We investigated how high a human could jump if muscle volume were optimized for jumping, and determined how the optimal parameters improve performance. We used a four-link inverted pendulum model of human vertical jumping actuated by Hill-type muscles, that well-approximates skilled human performance. We optimized muscle volume by allowing the cross-sectional area and muscle fiber optimum length to be changed for each muscle, while maintaining constant total muscle volume. We observed, perhaps surprisingly, that the reference model, based on human anthropometric data, is relatively good for vertical jumping; it achieves 90% of the jump height predicted by a model with muscles designed specifically for jumping. Alteration of cross-sectional areas-which determine the maximum force deliverable by the muscles-constitutes the majority of improvement to jump height. The optimal distribution results in large vastus, gastrocnemius and hamstrings muscles that deliver more work, while producing a kinematic pattern essentially identical to the reference model. Work output is increased by removing muscle from rectus femoris, which cannot do work on the skeleton given its moment arm at the hip and the joint excursions during push-off. The gluteus composes a disproportionate amount of muscle volume and jump height is improved by moving it to other muscles. This approach represents a way to test hypotheses about optimal human functional morphology. Future studies may extend this approach to address other morphological questions in ethological tasks such as locomotion, and feature other sets of parameters such as properties of the skeletal segments

Relationships between Clinician-Level Attributes and Fidelity-Consistent and Fidelity-Inconsistent Modifications to an Evidence-Based Psychotherapy

Background: Clinicians often modify evidence-based psychotherapies (EBPs) when delivering them in routine care settings. There has been little study of factors associated with or implications of modifications to EBP protocols. This paper differentiates between fidelity-consistent and fidelity-inconsistent modifications and it examines the potential influence of two clinician characteristics, training outcomes, and attitudes toward EBPs on fidelity-consistent and fidelity-inconsistent modifications of cognitive behavioral therapy in a sample of clinicians who had been trained to deliver these treatments for children or adults. Methods: Survey and coded interview data collected 2 years after completion of training programs in cognitive behavioral therapy were used to examine associations between successful or unsuccessful completion of training, clinician attitudes, and modifications. Modifications endorsed by clinicians were categorized as fidelity-consistent or fidelity-inconsistent and entered as outcomes into separate regression models, with training success and attitudes entered as independent variables. Results: Successful completion of a training program was associated with subsequent fidelity-inconsistent modifications but not fidelity-consistent modifications. Therapists who reported greater openness to using EBPs prior to training reported more fidelity-consistent modifications at follow-up, and those who reported greater willingness to adopt EBPs if they found them appealing were more likely to make fidelity-inconsistent modifications. Conclusions: Implications of these findings for training, implementation, EBP sustainment, and future studies are discussed. Research on contextual and protocol-related factors that may impact decisions to modify EBPs will be an important future direction of study to complement to this research

Union Administrative Practices: A Comparative Analysis

In response to growing challenges, many labor organizations are reevaluating themselves in an effort to become more efficient and effective. Their efforts, however, are limited by their frames of reference. Seldom do unions compare practices across labor movements. To expand these frames of reference we compare union administrative practices in three countries: Canada, the United Kingdom, and the United States. Two specific areas of union administration are examined — human resource/personnel practices and strategic planning. Results from these countries are presented and analyzed to identify and explain similarities and differences

Kinase insert domain receptor/vascular endothelial growth factor receptor 2 (KDR) genetic variation is associated with ovarian hyperstimulation syndrome

Background The objective of this investigation was to determine if kinase insert domain/vascular endothelial growth factor receptor 2 (KDR/VEGFR2) genetic variation was associated with the development of ovarian hyperstimulation syndrome (OHSS) in patients undergoing controlled ovarian hyperstimulation (COH). Methods This was a case-control study of 174 patients who underwent controlled ovarian stimulation. Patient blood samples were genotyped for single nucleotide polymorphisms (SNPs) spanning the KDR locus. OHSS development, clinical outcome variables, SNP and haplotype frequencies were compared between control (n = 155) and OHSS (n = 19) groups. Results Patients who developed OHSS had significantly higher response markers (estradiol levels of the day of hCG administration, number of follicles developed, number of eggs retrieved) than control patients. When adjusted for age and self-identified race, the rs2305945 G/T genotype was associated (P = 0.027) with a decreased risk (OR = 0.30; 95% CI = 0.10, 0.93) of developing OHSS using an overdominant model. The rs2305945 G/T variant was also associated with decreased COH response (number of follicles, number of eggs retrieved) in an overdominant model. The rs2305948, rs1870378, rs2305945 (C-T-G) haplotype was associated with both decreased COH response and OHSS risk (unadjusted OR = 0.10; 95% CI = 0.01, 0.80, P = 0.031). Conclusions The KDR receptor is believed to play a central role OHSS development and is a target for pharmacological prevention of OHSS. These results indicate that genetic variation in the KDR gene may impact individual risk of developing OHSS from COH. In addition, the rs2305948 SNP and C-T-G haplotype might serve as potential biomarkers for poor ovarian response to COH