ASAP: A pharmacy-level intervention to increase nonprescription syringe sales to reduce bloodborne illnesses.

BACKGROUND: Pharmacy syringe sales are effective structural interventions to reduce bloodborne illnesses in populations, and are legal in all but two states. Yet evidence indicates reduced syringe sales in recent years. This study was designed as a feasibility test of an intervention to promote syringe sales by pharmacies in Arizona. METHODS: A four-month pilot among three Arizona pharmacies measured feasibility and acceptability through monthly surveys to 18 enrolled pharmacy staff members. RESULTS: Pharmacy staff reported increased ease of dispensing syringes across the study. Rankings of syringe dispensing as 'easiest' among 6 measured pharmacy practices increased from 38.9 % at baseline to 50.1 % post intervention module training, and to 83.3 % at pilot conclusion. The majority (72.2 %) of pharmacy staff agreed that intervention materials were easy to use. Over 70 % indicated that the intervention was influential in their "being more open to selling syringes without a prescription to someone who might use them for illicit drug use," and 61.1 % reported that in the future, they were highly likely to dispense syringes to customers who would use them to inject drugs. A vast majority (92 %) reported being likely to dispense subsidized naloxone if available to their pharmacy at no cost. CONCLUSIONS: An education-based intervention was found to be feasible and acceptable to pharmacy staff and had an observed impact on perceptions of ease and likelihood of dispensing syringes without a prescription to people who may use them to inject drugs.

Characterization of Quasi-Keplerian, Differentially Rotating, Free-Boundary Laboratory Plasmas.

We present results from pulsed-power driven differentially rotating plasma experiments designed to simulate physics relevant to astrophysical disks and jets. In these experiments, angular momentum is injected by the ram pressure of the ablation flows from a wire array Z pinch. In contrast to previous liquid metal and plasma experiments, rotation is not driven by boundary forces. Axial pressure gradients launch a rotating plasma jet upward, which is confined by a combination of ram, thermal, and magnetic pressure of a surrounding plasma halo. The jet has subsonic rotation, with a maximum rotation velocity 23±3 km/s. The rotational velocity profile is quasi-Keplerian with a positive Rayleigh discriminant κ^{2}∝r^{-2.8±0.8} rad^{2}/s^{2}. The plasma completes 0.5-2 full rotations in the experimental time frame (∼150 ns).

Perpendicular Subcritical Shock Structure in a Collisional Plasma Experiment.

We present a study of perpendicular subcritical shocks in a collisional laboratory plasma. Shocks are produced by placing obstacles into the supermagnetosonic outflow from an inverse wire array z pinch. We demonstrate the existence of subcritical shocks in this regime and find that secondary shocks form in the downstream. Detailed measurements of the subcritical shock structure confirm the absence of a hydrodynamic jump. We calculate the classical (Spitzer) resistive diffusion length and show that it is approximately equal to the shock width. We measure little heating across the shock (<10% of the ion kinetic energy) which is consistent with an absence of viscous dissipation.

Time-resolved velocity and ion sound speed measurements from simultaneous bow shock imaging and inductive probe measurements.

We present a technique to measure the time-resolved velocity and ion sound speed in magnetized, supersonic high-energy-density plasmas. We place an inductive ("b-dot") probe in a supersonic pulsed-power-driven plasma flow and measure the magnetic field advected by the plasma. As the magnetic Reynolds number is large (RM > 10), the plasma flow advects a magnetic field proportional to the current at the load. This enables us to estimate the flow velocity as a function of time from the delay between the current at the load and the signal at the probe. The supersonic flow also generates a hydrodynamic bow shock around the probe, the structure of which depends on the upstream sonic Mach number. By imaging the shock around the probe with a Mach-Zehnder interferometer, we determine the upstream Mach number from the shock Mach angle, which we then use to determine the ion sound speed from the known upstream velocity. We use the sound speed to infer the value of Z̄Te, where Z̄ is the average ionization and Te is the electron temperature. We use this diagnostic to measure the time-resolved velocity and sound speed of a supersonic (MS ∼ 8), super-Alfvénic (MA ∼ 2) aluminum plasma generated during the ablation stage of an exploding wire array on the Magpie generator (1.4 MA, 250 ns). The velocity and Z̄Te measurements agree well with the optical Thompson scattering measurements reported in the literature and with 3D resistive magnetohydrodynamic simulations in GORGON.

An imaging refractometer for density fluctuation measurements in high energy density plasmas.

We report on a recently developed laser-probing diagnostic, which allows direct measurements of ray-deflection angles in one axis while retaining imaging capabilities in the other axis. This allows us to measure the spectrum of angular deflections from a laser beam, which passes through a turbulent high-energy-density plasma. This spectrum contains information about the density fluctuations within the plasma, which deflect the probing laser over a range of angles. We create synthetic diagnostics using ray-tracing to compare this new diagnostic with standard shadowgraphy and schlieren imaging approaches, which demonstrates the enhanced sensitivity of this new diagnostic over standard techniques. We present experimental data from turbulence behind a reverse shock in a plasma and demonstrate that this technique can measure angular deflections between 0.06 and 34 mrad, corresponding to a dynamic range of over 500.

Collective optical Thomson scattering in pulsed-power driven high energy density physics experiments (invited).

Optical collective Thomson scattering (TS) is used to diagnose magnetized high energy density physics experiments at the Magpie pulsed-power generator at Imperial College London. The system uses an amplified pulse from the second harmonic of a Nd:YAG laser (3 J, 8 ns, 532 nm) to probe a wide diversity of high-temperature plasma objects, with densities in the range of 1017-1019 cm-3 and temperatures between 10 eV and a few keV. The scattered light is collected from 100 µm-scale volumes within the plasmas, which are imaged onto optical fiber arrays. Multiple collection systems observe these volumes from different directions, providing simultaneous probing with different scattering K-vectors (and different associated α-parameters, typically in the range of 0.5-3), allowing independent measurements of separate velocity components of the bulk plasma flow. The fiber arrays are coupled to an imaging spectrometer with a gated intensified charge coupled device. The spectrometer is configured to view the ion-acoustic waves of the collective Thomson scattered spectrum. Fits to the spectra with the theoretical spectral density function S(K, ω) yield measurements of the local plasma temperatures and velocities. Fitting is constrained by independent measurements of the electron density from laser interferometry and the corresponding spectra for different scattering vectors. This TS diagnostic has been successfully implemented on a wide range of experiments, revealing temperature and flow velocity transitions across magnetized shocks, inside rotating plasma jets and imploding wire arrays, as well as providing direct measurements of drift velocities inside a magnetic reconnection current sheet.

Planning for chemical incidents by implementing a Delphi based consensus study.

This paper provides a practical approach to the difficulties surrounding planning for chemical incidents, based upon the results of a Delphi based consensus study. It is intended to offer advice, which can be implemented at regional and local prehospital and hospital level. The phases of the response that are covered include preparation, management of the incident, delivery of medical support during the incident, and recovery and support after the incident.

Delphi based consensus study into planning for chemical incidents.

OBJECTIVE: To achieve consensus in all phases of chemical incident planning and response. DESIGN: A three round Delphi study was conducted using a panel of 39 experts from specialties involved in the management of chemical incidents. Areas that did not reach consensus in the Delphi study were presented as synopsis statements for discussion in four syndicate groups at a conference hosted by the Department of Health Emergency Planning Co-ordination Unit. RESULTS: A total of 183 of 322 statements had reached consensus upon completion of the Delphi study. This represented 56.8% of the total number of statements. Of these, 148 reached consensus at >94% and 35 reached consensus at >89%. The results of the process are presented as a series of synopsis consensus statements that cover all phases of chemical incident planning and response. CONCLUSIONS: The use of a Delphi study and subsequent syndicate group discussions achieved consensus in aspects of all phases of chemical incident planning and response that can be translated into practical guidance for use at regional prehospital and hospital level. Additionally, areas of non-consensus have been identified where further work is required.

A humanized monoclonal antibody produced in transgenic plants for immunoprotection of the vagina against genital herpes.

The ability to produce monoclonal antibodies (Mabs) in plants offers the opportunity for the development of an inexpensive method of mucosal immunoprotection against sexually transmitted diseases. To investigate the suitability of plant-expressed Mabs for vaginal preventive applications, we compared a humanized anti-herpes simplex virus 2 (HSV-2) Mab expressed in mammalian cell culture with the same antibody expressed in soybean. We found these Mabs to be similar in their stability in human semen and cervical mucus over 24 h, their ability to diffuse in human cervical mucus, and their efficacy for prevention of vaginal HSV-2 infection in the mouse.

Soybeans transformed with a fungal phytase gene improve phosphorus availability for broilers.

Male broilers (n = 416) were used to compare the efficacy of providing dietary phytase either as a commercial supplement or as a recombinant protein in transformed soybean. From 7 to 21 d of age, broilers were fed a basal diet containing 0.20% nonphytate P (nP) with additional supplementation by fungal phytase as Natuphos or as raw transformed soybeans expressing recombinant phytase at 400, 800, or 1,200 U/kg. For comparison, broilers were also fed the basal diet containing 0.08, 0.16, or 0.24 added nP. The basal diet was fed as the negative control. Diets were consumed ad libitum as a mash. All excreta were collected from each pen from 18 through 20 d of age, and the birds were killed at 21 d of age. Supplementing the basal diet with nP linearly increased body weight gain, feed efficiency, feed intake, toe ash weight and percentage, and tibia shear force and energy. Phosphorus digestibility decreased linearly as nP level increased, but P excretion increased. Dietary phytase linearly increased growth rate, feed intake, toe ash weight and percentage, tibia shear force and energy, and P digestibility, whereas excretion was decreased. Except for P digestibility, there was no difference in efficacy of responses for performance, bone mineralization, and P excretion between the two sources of phytase. It appears from this study that phytase can improve growth performance of broilers fed low nP diets when provided either as a commercial supplement or in the form of transformed seeds.

Are physiological effects of sleep deprivation in the rat mediated by bacterial invasion?

Recent reports have indicated that rats subjected to total sleep deprivation (TSD) by the disk-over-water method and sacrificed when death appeared imminent showed aerobic bacteria in their blood. Yoked control rats did not. Extrapolating from these results, it has been suggested that the late body temperature declines and eventual deaths of TSD rats are caused by septicemia, and that other, earlier-appearing effects of TSD-including weight loss, increased energy expenditure, and regulation of temperature at a higher level-might be mediated by impaired host defenses against bacterial invasion. Three measures of aerobic bacterial invasion were used to evaluate these hypotheses: bacteremia, bacterial colonization in major organs of filtration (liver, kidney, and mesenteric lymph nodes), and adherence of bacteria to the cecal wall. Experiment 1 showed nonsignificant trends toward more bacterial invasion in 4-day TSD rats compared to yoked control rats and no relationship between the bacterial indicators and the early TSD effects. Experiment 2 showed that the elimination of aerobic bacterial infection by antibiotic treatment did not prevent the early TSD effects in 4-day TSD rats. Experiment 3 showed that the elimination of aerobic bacterial invasion in TSD rats did not eliminate the late temperature decline or the progression towards death. The results showed no significant evidence of aerobic bacterial invasion early in TSD and no indication that the major effects of TSD were dependent upon aerobic bacterial invasion.

Mutational analysis of a putative NTP-binding domain in the replication-associated protein (AC1) of bean golden mosaic geminivirus.

Bean golden mosaic virus (BGMV) is a whitefly-transmitted, ssDNA geminivirus with a bipartite genome. AC1 is the only ORF required for geminiviral replication. A putative NTP-binding motif, EGX4GKTX32DD, was present in the derived amino acid sequence of the replication-associated protein from the AC1 ORF for 13 geminiviruses including BGMV-GA (Guatemalan isolate, amino acids 221-263). We analyzed the phenotypes of mutations within this domain using a rapid and sensitive PCR-based assay for geminiviral replication developed for these studies. Replication in tobacco cells (NT-1 suspension cells) and infection of beans were abolished when codons were changed from K228 to H or D262 to R within the putative NTP-binding site. A temperature-sensitive replication phenotype was conferred by changing E221 to R within the putative NTP-binding domain. Replication was unaffected by changing a nonconserved codon near the putative NTP-binding domain from 1190 to R. Our results demonstrate that the putative NTP-binding domain is required for geminiviral replication. The role of NTP hydrolysis and the possible value of these mutants in a trans-dominant interference scheme for virus-derived resistance are discussed.

Stable transformation of Phaseolus vulgaris via electric-discharge mediated particle acceleration.

Transgenic Phaseolus vulgaris or common bean has been produced using electric-discharge particle acceleration. The method uses particle acceleration to introduce DNA into bean seed meristems. Multiple shoots are then generated and screened to recover transgenic plants at a rate of 0.03% germline transformed plants/shoot. We have been able to recover transgenic plants using both GUS and herbicide screening to introduce the gus, bar, and bean golden mosaic virus coat protein genes into the navy bean cultivar, Seafarer. The transgenic plants have been characterized over 5 generations of self-fertilization with no loss of introduced genes or expression. In addition, several families have been crossed with non-transgenic parents and these plants also show expected inheritance patterns. The introduced bar gene has been shown to confer strong resistance in transgenic beans to basta herbicide application in the greenhouse.

Use of the asymmetric polymerase chain reaction and DNA sequencing to determine genetic variability of bean golden mosaic geminivirus in the Dominican Republic.

A combination of the polymerase chain reaction (PCR), asymmetric PCR (A-PCR) and DNA sequencing was used to determine the nucleotide sequence of a hypervariable region of the bipartite genome of bean golden mosaic geminivirus (BGMV). This region, which was part of the intergenic region of the DNA-B component, was amplified using primers designed from the nucleotide sequence of a DNA-B component clone (pDRB1) of an isolate of BGMV from the Dominican Republic (BGMV-DR). pDRB1 is infectious on beans when coinoculated with the DNA-A component of BGMV-DR (pDRA1), and typical bean golden mosaic symptoms are observed on infected plants. Bean leaf tissue infected with BGMV was collected at five separate field locations in the Dominican Republic and the hypervariable region was amplified by PCR, ssDNA was produced using A-PCR, and partial nucleotide sequences were determined. The sequences of the hypervariable region from the field-collected samples ranged from 95% (one sample) to 98% (four samples) identical to the sequence of pDRB1. This contrasts with sequence identities of 86, 75 and 46% between the pDRB1 hypervariable region and the hypervariable regions of BGMV isolates from Guatemala, Puerto Rico and Brazil respectively, and 42% with bean dwarf mosaic geminivirus. These results indicate that Dominican Republic isolates of BGMV are very similar and should be considered isolates of the same virus (BGMV-DR), and that the infectious clones of BGMV-DR are representative of BGMV isolates in the Dominican Republic. The procedures described for DNA extraction from leaf tissue and for production of high quality ssDNA using PCR and A-PCR are rapid and efficient and could be applied to studies of variability and epidemiology of other viruses.

Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration.

Three different target tissues (protoplast-derived cells, nodules, and stems) and two unrelated hybrid genotypes of Populus (P. alba x P. grandidentata 'Crandon' and P. nigra 'Betulifolia' x P. trichocarpa) have been stably transformed by electric discharge particle acceleration using a 18.7 kb plasmid containing NOS-NPT, CaMV 35S-GUS, and CaMV 35S-BT. Four transformed plants of one hybrid genotype, NC5339, containing all 3 genes were recovered and analyzed. Two expressed GUS and one was highly resistant to feeding by 2 lepidopteran pests (the forest tent caterpillar, Malacosoma disstria, and the gypsy moth, Lymantria dispar.) Pretreatment of the target tissues, fine-tuning of the bombardment parameters, and the use of a selection technique employing flooding of the target tissues were important for reliable recovery of transformed plants.

In vitro characterization of hybrid promoters and altered tryptophan operon promoters.

This study examines the in vitro interaction of hybrid and altered Escherichia coli promoters and other promoters with purified E. coli RNA polymerase. Three parameters of polymerase activity were examined: the time for open complex formation; the temperature of transitions; and the time required for productive initiation. The results indicate the rate of in vitro binding as measured by the filter binding technique does not completely correlate with the in vivo activities among these diverse promoters. Transition temperatures ranged from 13 to 27 degrees C with the lowest transition temperatures associated with the relatively weak in vivo beta-lactamase and anti-tet promoters. The productive initiation studies showed a dependence on labeled nucleoside triphosphate concentration when that nucleotide was present early and frequently in the transcript. Promoters containing the -10 region of the lac promoter had slow productive initiation rates while trp -10 promoter derivatives were generally very fast. In the promoters studied here, a trend was noted between the binding rate and transition temperature studies in that the promoters with the lower transition temperatures tended to bind more rapidly.

Role of DNA regions flanking the tryptophan promoter of Escherichia coli. I. Insertion of synthetic oligonucleotides.

To examine the effect of altering the nucleotide sequence near the promoter on its activity, pKO-1 vector derivatives have been constructed which allow insertion of DNA fragments at specified sites upstream or downstream from the trp promoter. Oligonucleotides that might be expected to alter the melting properties, or have a tendency to form a distinctive nonstandard structure were introduced. These oligonucleotides had the repeating dinucleotide sequences GC, AT or AG. Sequence analysis of the inserts and studies of the relative galactokinase expression from the altered plasmids indicated that changes upstream from the trp promoter at -39 or beyond had little effect on trp promoter activity, whereas changes at +2 or farther downstream produced up to two-fold increases in gene expression, as compared to the control plasmid.