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The European Bioanalysis Forum is a bioanalytical nonprofit organization comprised of European pharmaceutical companies (27 members to date) and currently expanding to include CROs as well. The European Bioanalysis Forum provides a broad European bioanalytical network for the discussion of scientific, technological and regulatory topics of bioanalytical interest. The 3rd Annual Open Symposium was again much anticipated after the two previous successful meetings. The symposium included sessions on thinking outside the ‘commodity’ box, bioanalytical challenges with blood, global harmonization, assay platforms, dried blood spots, immunogenicity, matrix effects, anomalous results, biomarkers and two plenary technology sessions hosted by the Platinum sponsors. Experts and key opinion leaders were invited as guest speakers. A total of 424 delegates registered from 113 companies representing a large percentage of the European bioanalytical community. In addition to 48 oral presentations, 88 posters were presented and there was a vendor exposition of 40 companies.

The European Bioanalysis Forum is a non-profit organization comprised of European pharmaceutical companies (25 members to date). Their activities focus on bringing together managers and scientists in the broad field of bioanalysis to discuss topics related to science, process and regulations. There has been much interest over the past few years in the potential application of dried blood spots as an alternative to traditional plasma collection in pharmacokinetic studies. The success of the technique has been highlighted by several companies. We know that seven of the European Bioanalysis Forum member companies are using dried blood spots intensively and that 22 out of 25 companies are using it or plan to use it very soon, initially in nonregulated studies. However, most companies have less than 1 year of experience with dried blood spots and, beyond the scientific merit, it is less clear just how the technique is perceived by key client groups, such as toxicology, clinical and regulatory authorities. The objective of this symposium was to bring together representatives from all of these client groups as well as a broad bioanalytical audience to discuss the various perspectives on dried blood spots and to try to provide answers to potential issues that still remain. The symposium included sessions on dried blood spots in the nonregulated environment, toxicology and regulatory/QA perspectives, clinical use and bioanalytical applications and tools. There was plenty of time for discussion within the program in tutorials, poster and break-out sessions and the degree of delegate participation reflected the high level of engagement with the topic. A total of 190 delegates attended from more than 80 organizations. In addition to 21 oral presentations, more than 20 posters were presented and there was a vendor exposition of ten sponsor companies.

UNLABELLED PREMISE OF THE STUDY Primers were developed for a portion of the ycf1 plastid gene in magnoliid taxa to investigate the utility of ycf1 in phylogenetic analyses. • METHODS AND RESULTS Twenty-six species across six families within the magnoliid group (Canellales, Piperales, Laurales, and Magnoliales) were sampled to examine the ability to amplify ycf1. Additionally, 29 accessions of Asimina and Deeringothamnus (Annonaceae) were sequenced to assess levels of variation in ycf1 compared to matK and trnL-F. • CONCLUSIONS Results indicate that ycf1 is easily amplified and sequenced. In Annonaceae, ycf1 provides more informative phylogenetic characters than commonly used markers such as matK and trnL-F.

The European Bioanalysis Forum is a bioanalytical discussion group comprised of European pharmaceutical companies (27 members to date). The membership share a common vision to advance the shared understanding of topical concerns through discussion of scientific, technological and regulatory issues of bioanalytical interest. The 2nd Annual Open Symposium was much anticipated after the success of the first one. The symposium included sessions on the European Medicines Agency draft bioanalytical guidance, challenges in the analysis and validation of biomarkers, metabolite quantification issues, dried blood spot technology, the analysis of proteins and peptides by MS, latest hot topics in ligand-binding assays, unresolved issues in method validation and a plenary session from the Platinum sponsors. Experts and key opinion leaders were invited as guest speakers. A total of 420 delegates (up from 251 delegates in 2008) attended from 160 organizations, representing a large percentage of the European and international bioanalytical community. In addition to 40 oral presentations, 72 posters were presented and there was a vendor exposition of 35 companies.

Following intensive discussions, review, alignment of procedures and multiple surveys among their member companies, the European Bioanalysis Forum (EBF) is providing a recommendation on how to integrate incurred sample reproducibility (ISR) in the bioanalytical process. The recommendation aims to provide guidance throughout the lifecycle of a validated method, including the application of the method in study support. In its recommendation, the EBF considers both the internal discussions with EBF member companies, as well as the input provided in international meetings where ISR was discussed. The ultimate goal of the EBF recommendation is to ensure that bioanalytical methods can provide accurate and reproducible concentration data for pharmacokinetic and/or toxicokinetic evaluation, without any compromise, while safeguarding the optimal use of laboratory resources.

Pachyanthus (Melastomataceae: Miconieae) is traditionally circumscribed as a group of 20 species of Caribbean shrubs, mostly local Cuban endemics. However, the exact number of species is unknown because generic delimitations are still unclear for most genera of Miconieae. To test the monophyly of Pachyanthus, we produced a molecular phylogeny using nrITS and plastid trnS-G sequence data for 38 taxa of Miconieae in the genera Pachyanthus, Calycogonium, Tetrazygia, and Miconia. A species of Graffenrieda, a member of the sister group, Merianieae, was used as an outgroup. In the single-gene and combined analyses Pachyanthus is never recovered as a monophyletic group in any of its traditional circumscriptions. However, it is possible to distinguish a clade, Pachyanthus s.s., which is characterized by ovate to obovate petals that are conspicuously unguiculate; calyx lobes connate before anthesis, that separate into sepal-like segments; straight, ovate to oblong, yellow anthers without connective appendages or, if present, then inconspicuous, anthers dehiscing by small apical pores, and fruits with numerous seeds, 1–2 mm long, obpyramidal, with a smooth testa. The remaining species traditionally included in Pachyanthus do not have any of these features. They do not form a single monophyletic group, but instead are scattered across several clades, which also contain species of Calycogonium, Miconia, and Tetrazygia. Additional detailed molecular and morphological studies are needed in the Miconieae to ascertain whether these clades should be treated as new genera or recircumscribed as part of currently recognized genera in the Caribbean region, such as Calycogonium and Tetrazygia.

As traditionally circumscribed, the genus Polygala has been shown to be non-monophyletic, and most of the necessary generic combinations have been made for the New World species. The genus Hebecarpa, though, still needs to be addressed, as only eight species have been transferred. Even though over 65 names within Polygala are referable to Hebecarpa, preliminary study indicates that more than half the names are likely not worthy of taxonomic recognition, but careful revisionary study is necessary for fully sorting out the species and accurate synonymisation. To make the most accurate names available for modern workers, we here transfer 11 names (and associated synonyms) which we think most likely reflect discreet evolutionary lineages (species), bringing the total number of Hebecarpa species to 19. Forty-one additional names in Polygala that are attributable to Hebecarpa are enumerated but left for synonymisation or transfer after more study.

In bioanalysis of small molecules, the analyte concentration in the measured samples should reflect the concentration during sample collection. Precautions may be needed to prevent over- or under-estimation of the obtained result. This might require the addition of stabilizers to prevent degradation or nonspecific binding. For unstable drugs, it is essential to know how analytes can be stabilized before the start of the clinical study. Although the stabilization methods are well documented, the impact of the stabilization on the clinical workflow is not properly addressed. Already during method development, the clinical implications in terms of personnel safety, ease of use, training possibilities and staff capacity should be taken into account, and validation of the bioanalytical method should reflect collection procedures.

The commercial harvest of American matsutake (Tricholoma magnivelare) is a multi-million dollar industry in the Pacific Northwest region of NorthAmerica. Howtobestmanageforsustainable mushroomproductionisuncertainandconcerns remainaboutthecumulativeeffectsofpicking in the same areas year-after-year and whether raking of surface litter and mineral soil layers to find mushrooms will reduce subsequent fruiting. Here, we evaluate the effects of several mushroom harvest techniques on American matsutake production. This study was established in the Oregon Cascades in 1994 with the selection of 18 shiros of similar mushroom production. Six mushroom harvest treatments were implemented in 1995: (1) control, (2) best management practice (BMP), (3) shallow rake, litter replaced, (4) shallow rake, no replacement, (5) deep rake, litter replaced, (6) deep rake, no replacement. These treatments were pooled into three litter disturbance groups for analysis: (a) no raking of the litter, (b) litter raked with replacement, and (c) litter raked without replacement. Matsutake production on additional shiros was monitored to further compare the control and BMP treatments. Our results demonstrate that careful picking (BMP) was not detrimental to mushroom production during the initial 10 years of mushroom harvest activity. One-time treatments in which the forest floor litter layers were removed and not replaced were strongly detrimental to matsutake production and the effects have persisted for 9 years. Matsutake production was reduced to an intermediate degree by the raking with litter replacement treatments. Damage to shiros caused by repeated raking was not tested, however we expect that the effects of repeated raking would be more severe than those reported here. Negative treatment effects were particularly noticeable in years with abundant fruiting. When environmental conditions are poor for fruiting all shiros experience low production, thereby obscuring treatment effects. Within-year and year-to-year variation in fruiting is a major challenge to studies of matsutake ecology, particularly with regard to documenting treatmenteffects. Furtherstudiesspanningyearsorevendecades willlikelybeneededtoquantifyproduction,effectsofmanagementactivities,and investigate the biology of Tricholoma magnivelare. Because this study was limited to one habitat type, extension of the results to substantially different habitats types must be made with caution. However, we speculate that since the underlying biology of matsutake fruiting is similar across a wide range of habitats, careful picking should generally not hinder subsequent fruiting when other substantial disturbance to the shiro is absent. # 2006 Elsevier B.V. All rights reserved.

Aim To assess the pharmacokinetic effect and tolerability of lamotrigine 200 mg day−1 and olanzapine 15 mg day−1 coadministration in healthy male volunteers. Methods Subjects were randomized to receive either lamotrigine titrated on days 1–42 with olanzapine added on days 43–56 (LTG + OLZ group; N = 16), lamotrigine titration with placebo added on days 43–56 (LTG group; N = 12), or placebo on days 1–42 with olanzapine added on days 43–56 (OLZ group; N = 16). Steady state (0–24 h) pharmacokinetic profiles were determined on day 56 in each group. Results The average (90% confidence interval) ratios of lamotrigine area under the concentration–time curve from 0 to 24 h and maximum observed concentration for the comparison of LTG + OLZ:LTG were 0.76 (0.65, 0.90) and 0.80 (0.71, 0.90), respectively. Olanzapine pharmacokinetics were essentially unaffected by lamotrigine. The most frequently reported adverse events (AEs) during combination therapy were fatigue, dizziness and mild transaminase elevations. These AEs occurred at similar frequencies in the LTG + OLZ and OLZ cohorts, while being less frequent or absent in the LTG group. Conclusions Lamotrigine and olanzapine coadministration in patients who may benefit from the combination was supported by this study. Lamotrigine dosing schedules are recommended to remain unchanged when combined with olanzapine therapy. However, the possibility exists that the lamotrigine dose for some patients may need adjustment to optimize treatment when olanzapine is added to or withdrawn from a regimen including lamotrigine.

The vascular flora of an abandoned limestone quarry adjacent to the Kentucky River in Clark County, Kentucky, was studied from 1993 through 1999. A total of 260 species in 181 genera from 64 families were found in five diverse habitats. Fifty-seven (21.9%) were Clark County distributional records. Seventy-three (28.1%) were exotic species; 22 of these are Kentucky naturalized invasive pest plants. Taxa are classified into Equisetophyta (2), Polypodiophyta (2), Pinophyta (1), and Magnoliophyta (255). Largest families were Asteraceae (43), Poaceae (30), Fabaceae (15), Cyperaceae (13), and Rosaceae (10). Two state-listed species, Liparis loeselii and Spiranthes lucida, were present in the quarry.

The measured performance of analog circuits is at some level impacted by a broad array of unintended real-world parameters in the form of: printed circuit board parasitic, component value variations, and non-idealities associated with cabling and connectors. simulation of these types of systems can rapidly become computationally intractable when more variables are considered. In this paper, analytic methods are developed and compared to measured data in order to predict the impact of parasitic on the performance of a switching power supply output filter. The filter is first described using state space methods, and then converted to a standard linear singularly perturbed system for the inclusion of parasitic parameters. The techniques presented in this paper are readily scalable to more complicated electronic circuits yielding computationally compact methods to accurately predict system performance.

Accelerator mass spectrometry (AMS) is being used more widely to provide PK data for early decision making or to generate absolute bioavailability data in later phases of development. Presently, there is no clear consensus on the level of the scientific validation required for these assays. The European Bioanalysis Forum (EBF) has conducted two surveys with its members and presented the results at its 4th Open Symposium. With AMS being used for discrete scientific assessment, method establishment of AMS assays should focus on science rather than trying to fit the assay parameters into validation criteria used for Regulated Bioanalysis guidance, and an amount of freedom of execution and interpretation is needed. Hence, the EBF focuses their recommendation on introducing terminology around scientific qualification or validation to be used in relation to AMS. Guidance is given on which parameters should be investigated when a qualified method is required. The recommendations of the EBF for scientific validation are described herein. The scientific validation of AMS assays will be different to that applied for LC–MS/MS assays, and an example is that accuracy and precision limits, as used for ligand-binding assays, would be more appropriate.

Background: Direct analysis of dried blood spot (DBS) samples was investigated using a prototype semi-automated robotic device that allows the direct elution of sample spots from a DBS paper card to an online SPE cartridge. The eluted SPE samples were analyzed with high-performance LC–MS/MS. Results: A LLOQ of 0.01 ng/ml was achieved with a linear calibration range from 0.01 to 25 ng/ml. Optimal performance data were obtained from spotting the internal standard solution on the card before blood spotting. Internal standard addition from the system injector loop produced intra-assay inaccuracy of -9.0–7.3% and precision of 1.3–8.2%, and inter-assay inaccuracy of -3.5–3.9% and precision of 4.4–8.7%. Conclusion: Results demonstrated the feasibility of a semi-automated online rapid direct elution method that avoids manual extraction for DBS sample analysis using the online DBS-SPE system coupled to LC–MS/MS.

A two-layered polymeric membrane is employed for the formation of separated dried plasma spots from whole blood as an alternative to the direct analysis of whole dried blood spots (DBS). This dried plasma spot (DPS) analysis procedure precludes potential issues of hematocrit differences in whole blood samples while providing pharmokinetic data from plasma rather than whole blood. The described procedure is also semi-automated thus providing a simpler work flow for LC/MS/MS bioanalysis procedures.Molecular filtration of red blood cells (RBC) from applied microsamples of whole blood fortified with guanfacine and its stable isotope internal standard was accomplished with a two-layer polymeric membrane substrate. The lower membrane surface containing the separated plasma spot was physically separated from the upper membrane followed by semi-automated direct elution of the sample to an online solid-phase extraction (SPE) cartridge followed by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS).A two-layer membrane sample preparation substrate produced plasma from whole blood without centrifugation which could be directly eluted for semi-automated LC/MS/MS bioanalysis. Standard curves were constructed by plotting peak area ratios between the analyte and the stable isotope labeled internal standard (SIL-IS) versus the nominal concentration in whole blood. A weighted 1/x(2) linear regression was applied to the data from DPS samples. Standard curves were linear over the range 0.25-250 ng/mL human whole blood. The representative regression equation was y = 0.0142x + 0.00248 (R(2) = 0.995) for the described DPS assay.The described work demonstrates proof-of-principle using membrane sample preparation techniques to form DPS samples from whole blood for subsequent bioanalysis by LC/MS/MS. This approach has the potential to eliminate the hematocrit issues from the current controversy surrounding validation of DBS assays.

The two 4 km long gravitational wave detectors operated by the Laser Interferometer Gravitational-wave Observatory (LIGO) were modified in 2008 to read out the gravitational wave channel using the DC readout form of homodyne detection and to include an optical filter cavity at the output of the detector. As part of the upgrade to Enhanced LIGO, these modifications replaced the radio-frequency (RF) heterodyne system used previously. We describe the motivations for and the implementation of DC readout and the output mode cleaner in Enhanced LIGO. We present characterizations of the system, including measurements and models of the couplings of the noises from the laser source to the gravitational wave readout channel. We show that noise couplings using DC readout are improved over those for RF readout, and we find that the achieved shot-noise-limited sensitivity is consistent with modeled results.

The European Bioanalysis Forum (EBF) is a bioanalytical discussion group comprised of European pharmaceutical companies (26 members to date). The membership shares a common vision to advance the shared understanding of topical concerns through discussion of scientific, technological and regulatory issues of bioanalytical interest. The objective of the open symposium was to reach out to the broader European and global bioanalytical community, to report back on discussions EBF member companies have during their biannual closed meetings and, going forward, to provide guidance and recommendations to the European and global bioanalytical community. Pharmaceutical and biotechnology companies, contract research organisations (CROs), academia and instrument vendors were invited to attend to exchange scientific ideas and information on topical issues of mutual concern. The symposium included sessions on assay validation, metabolite quantification, instrumentation developments and protein quantification and immunogenicity. Experts and key opinion leaders were invited as guest speakers. A total of 251 delegates attended, representing a large percentage of the European bioanalytical community. The majority of attendees represented pharmaceutical companies (36%), CROs (42%), instrument vendors (15%) and academia (4%). In addition to 18 oral presentations, 51 posters were presented and there was a well-supported vendor exposition.

The Empire State Electric Energy Research Corporation (ESEERCO) contracted with ACRT, Inc. Environmental Specialist to research the status and efficacy of Tree Growth Regulators (TGR's) and their applicability for the electrical utility industry. The project included the preparation of an annotated bibliography, a TGR practitioner survey for usage data and unpublished reports, a synopsis of the bibliography, and an information dissemination/storyboard session for ESEERCO members and TGR scientists. The user survey had 160 respondents and their responses are tabulated and discussed. Various TGR application methods—trunk injection, bark banding, trunk implant, soil application—are discussed and evaluated. This presentation summarizes the user information and the pertinent results of the storyboard session.

To assess the pharmacokinetic effect and tolerability of lamotrigine 200 mg day(-1) and olanzapine 15 mg day(-1) coadministration in healthy male volunteers.Subjects were randomized to receive either lamotrigine titrated on days 1-42 with olanzapine added on days 43-56 (LTG + OLZ group; N = 16), lamotrigine titration with placebo added on days 43-56 (LTG group; N = 12), or placebo on days 1-42 with olanzapine added on days 43-56 (OLZ group; N = 16). Steady state (0-24 h) pharmacokinetic profiles were determined on day 56 in each group.The average (90% confidence interval) ratios of lamotrigine area under the concentration-time curve from 0 to 24 h and maximum observed concentration for the comparison of LTG + OLZ:LTG were 0.76 (0.65, 0.90) and 0.80 (0.71, 0.90), respectively. Olanzapine pharmacokinetics were essentially unaffected by lamotrigine. The most frequently reported adverse events (AEs) during combination therapy were fatigue, dizziness and mild transaminase elevations. These AEs occurred at similar frequencies in the LTG + OLZ and OLZ cohorts, while being less frequent or absent in the LTG group.Lamotrigine and olanzapine coadministration in patients who may benefit from the combination was supported by this study. Lamotrigine dosing schedules are recommended to remain unchanged when combined with olanzapine therapy. However, the possibility exists that the lamotrigine dose for some patients may need adjustment to optimize treatment when olanzapine is added to or withdrawn from a regimen including lamotrigine.

A 10-year survey of vascular plants was made of Rock Creek Research Natural Area, a 77-ha deep, narrow gorge of old-growth Hemlock-Mixed Mesophytic Forest located in Laurel County, Kentucky, on the Daniel Boone National Forest. The study documented 350 specific and infraspecific taxa in 223 genera and 93 families. Thirteen are nonindigenous naturalized species. Vascular plants include 6 Lycopodiophyta, 25 Polypodiopyta, 5 Pinophyta, and 314 Magnoliophyta; 255 are annual, biennial, and perennial herbs and 95 are woody vines, shrubs, and trees. Seventeen rare and special interest species have been recorded, including 4 that are listed by the state. The floristic survey provides a baseline reference for relative abundance, species richness, plant associations, habitats, and generalized life-forms within the Rock Creek Research Natural Area.

Non-communicating arachnoid cyst of the lateral temporal bone is a rare condition. We present a case of a non-communicating arachnoid cyst of the temporal bone in an infant who presented with a lower motor neuron facial nerve paralysis. The patient was treated by surgical excision of the cyst.

Background.Computers that collect data from patients and provide both patients and practitioners with printed feedback on a range of health risks are a tool for assisting general practitioners with preventive care. This study assessed the impact of computer-generated printed feedback on cervical screening among women who were underscreened for cervical cancer. Method.Female attenders at two Australian general practices were randomly allocated to Experimental or Control groups. Women in both groups completed a health risk survey on a touch screen computer prior to their consultation. Those in the Experimental group received printed pages summarizing their results, including their eligibility for cervical screening and last Pap test, for themselves and their doctor. The number and proportion of underscreened women who had a Pap test in the 6 months after completing the computer survey, as determined by pathology records, were examined. Results.Of the 679 participants, 139 were classified as underscreened on the basis of self-report (74 Experimental, 65 Control) and 272 on the basis of their pathology records (148 Experimental, 124 Control). Overall about one-third of women had a test in the 6-month period, and the differences between the groups were not significant for women overall (18–70 years) or for women 18–49 years. Among women 50–70 who were underscreened based on self-report, those receiving the printout were more likely to have a Pap test in the next 6 months (P< 0.05). This pattern was also evident, but did not reach statistical significance, for older women who were underscreened based on pathology records. Conclusions.We are unable to draw conclusions regarding the effectiveness of the computer system due to the modest proportions of women screened, the small numbers, and the fact that the computer survey may have created an intervention effect in the Control group. As the study suggests the computer system is acceptable to women and may be effective for encouraging screening among older women, further exploration of the system is desirable.

The diffusion behaviour of an aqueous mixture of the sugar-based non-ionic surfactant dodecylmalono-bis-N-methylglucamide DBNMG and the anionic surfactant of similar tail length, sodium dodecyl sulfate SDS, are examined in the presence of 5 mass% gelatin; the results clearly show that there are two types of mixed micelle present in the system arising through competition between gelatin and the non-ionic micelle for the anionic surfactant.

There are more than 46,000 tree and landscape service companies in the U.S.A. according to the most recent government census. The majority of these firms are good, experienced arborists/landscapers, but they have little or no formal business training. Assistance to Green Industry small businesses is available from numerous government agencies, educational organizations, commercial and trade associations, financial groups and business consultants. Some of these free or low-cost businesses management assistance resources are available to tree or landscape service companies wherever they are located, but they need to look for them. Tree and landscape business owners must truly "Become Business Professionals as well as Tree Professionals" in order to prosper and survive in today's economic and regulated society.

Utility line clearing has developed into a major industry. Many utilities today employ professionals (arborists, foresters, etc.), to direct their programs. Increased federal and state regulations have changed work procedures, tools, methods, equipment, etc. Basically the line clearance industry, with the exception of telephone/telegraph, has reacted to crisis. There is a need for a research coordinating committee to establish priorities, goals and ultimately stabilize or reduce maintenance line clearing cost.

Tree care, pruning and removal operations have been difficult to price because each tree, each location, and each situation is different. Traditionally, tree care salespeople have learned to estimate jobs by the trial-and-error method. Many times, the salesperson has not been in sales over a long enough period of time to gain pricing experience. The concept of pricing using fixed and variable factors for the different tasks can accommodate the differences in trees, locations, and situations. The system can, and should be modified to each company's operations. The use of man-hours required to perform specific tree care operations provides the key to calculating costs and also provides a measurable basis to evaluate crew performance.