Insight from Molecular, Pathological, and Immunohistochemical Studies on Cellular and Humoral Mechanisms Responsible for Vaccine-Induced Protection of Rainbow Trout against <em>Yersinia ruckeri</em>

The immunological mechanisms associated with protection of vaccinated rainbow trout, Oncorhynchus mykiss, against enteric redmouth disease (ERM), caused by Yersinia ruckeri, were previously elucidated by the use of gene expression methodology and immunochemical methods. That approach pointed indirectly to both humoral and cellular elements being involved in protection. The present study correlates the level of protection in rainbow trout to cellular reactions in spleen and head kidney and visualizes the processes by applying histopathological, immunohistochemical, and in situ hybridization techniques. It was shown that these cellular reactions, which were more prominent in spleen than in head kidney, were associated with the expression of immune-related genes, suggesting a Th2-like response. Y. ruckeri, as shown by in situ hybridization (ISH), was eliminated within a few days in vaccinated fish, whereas nonprotected fish still harbored bacteria for a week after infection. Vaccinated fish reestablished normal organ structure within a few days, whereas nonprotected fish showed abnormalities up to 1 month postinfection. Protection in the early phase of infection was mainly associated with the expression of genes encoding innate factors (complement factors, lysozyme, and acute phase proteins), but in the later phase of infection, increased expression of adaptive immune genes dominated. The histological approach used has shown that the cellular changes correlated with protection of vaccinated fish. They comprised transformation of resident cells into macrophage-like cells and increased occurrence of CD8α and IgM cells, suggesting these cells as main players in protection. Future studies should investigate the causality between these factors and protection

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Taxotere-, Zometa-, og palliativ strålebehandling hos pasienter med hormonrefraktær prostatakreft ved Ahus

Bakgrunn: Det har vært reist spørsmål om de pasienter som trenger det, får behandling med Taxotere, og andre palliative behandlingstiltak. Vi har derfor kartlagt bruken av Taxotere mht effekt og bivirkninger. Materiale og metode: En retrospektiv databasert studie (DIPS) er gjort på alle pasienter i kontakt med sykehuset i løpet av 2006 som hadde kombinasjonen av diagnosekode C61 (CaP) og en eller flere metastasekoder (n=69). Sekstien pasienter ble funnet (i 2006). Journaldata ble registrert ut 2007. Av disse pasientene plukket vi ut de som var i hormonrefraktær fase i løpet av observasjonstiden. Resultater: 48 pasienter (79%) kom i hormonrefraktær fase (HRF), definert som PSA stigning i tre påfølgende blodprøver etter initialt fall på hormonbehandling. Seks av disse hadde ikke symptomer ila observasjonstiden (på smertestillende behandling). Alle pasientene hadde skjelettmetastaser, og 22 (46%) hadde metastaser allerede på diagnosetidspunkt. Andelen Gleason 8-10 (hissig cancer) i materialet var 26 (54%). Andelene T1 var 8 (17%), T2 13 (27%), T3 26 (54%) og T4 1 (2%) (ved diagnosetidspunkt). Palliativ stålebehandling ble gitt til 31 (65%) av pasientene. Fem (10%) fikk palliativ stråling mot prostata, 7 (15%) fikk channelling TUR-P. Tretten (27%) fikk tilbud om Taxotere. Alle hadde symptomgivende sykdom. Fire fikk ikke behandling pga raskt progredierende sykdom. Ni (19%) pasienter fikk Taxotere. Ant.pas. (%) Alder (range) Total tid* n = 48 v/ sykd debut v/HRF i mnd til HRF Taxotere (+Zometa hos 7) 9 (19%) 63.1 (53-75) 67,6 (54-77) 20,4 (14-26) Kun Zometa 19 (40%) 69,0 (55-75) 72,3 (62-82) 19,2 (16-33) Annen pall.beh. 20 (41%) 67,3 (50-79) 71,8 (52-82) 20,4 (17-31) ----------------------------------------------------------------------------------------------------------------- Total: 48 (100%) 67,2 (50-79) 71,2 (52-82) 20,0 (=1 2/3år) *Gjennomsnittstiden er beregnet fra oppstart hormonbehandling med LHRH analog til PSA begynner å stige. Målt fra oppstart antiandrogenbehandling: 23 mnd, 2 år (range 17-33 mnd). I snitt gikk det 7 måneder (3-11 mnd) mellom PSA stigning (”HRF”) og oppstart av Taxotere. Fire av 9 (44%) avsluttet behandlingen etter 2-6 beh pga bivirkninger (3 fikk neutropen feber, 1 pga GI-plager og tretthet). Fem av 9 (56%) avslutter behandlingen pga biokjemisk sykdoms-progresjon etter 7-24 behandlinger, i snitt 8 mnd (4-12 mnd) etter oppstart Taxotere. Med unntak av de som fikk neutropen feber, så vi behandlingseffekt på PSA hos alle som fikk Taxotere over noe tid. Vi fant reduksjon i opiatforbruket hos 4/9 (44%) på Taxotere, og 12/19 (63%) på Zometa. Konklusjon: Taxotere tilbys circa ¼ av pasientene, og kun circa halvparten tåler behandlingen. Palliativ strålebehandling gis i stor utstrekning. Taxotere-pasientene tenderer til å være yngre enn de som får Zometa. Ahus synes å være aktive med hensyn til palliativ, resurskrevende behandling for CaP pasienter, men Taxotere gis sent i forløpet

TASK 3.4--IMPACTS OF COFIRING BIOMASS WITH FOSSIL FUELS

With a major worldwide effort now ongoing to reduce greenhouse gas emissions, cofiring of renewable biomass fuels at conventional coal-fired utilities is seen as one of the lower-cost options to achieve such reductions. The Energy & Environmental Research Center has undertaken a fundamental study to address the viability of cofiring biomass with coal in a pulverized coal (pc)-fired boiler for power production. Wheat straw, alfalfa stems, and hybrid poplar were selected as candidate biomass materials for blending at a 20 wt% level with an Illinois bituminous coal and an Absaloka subbituminous coal. The biomass materials were found to be easily processed by shredding and pulverizing to a size suitable for cofiring with pc in a bench-scale downfired furnace. A literature investigation was undertaken on mineral uptake and storage by plants considered for biomass cofiring in order to understand the modes of occurrence of inorganic elements in plant matter. Sixteen essential elements, C, H, O, N, P, K, Ca, Mg, S, Zn, Cu, Fe, Mn, B, Mo, and Cl, are found throughout plants. The predominant inorganic elements are K and Ca, which are essential to the function of all plant cells and will, therefore, be evenly distributed throughout the nonreproductive, aerial portions of herbaceous biomass. Some inorganic constituents, e.g., N, P, Ca, and Cl, are organically associated and incorporated into the structure of the plant. Cell vacuoles are the repository for excess ions in the plant. Minerals deposited in these ubiquitous organelles are expected to be most easily leached from dry material. Other elements may not have specific functions within the plant, but are nevertheless absorbed and fill a need, such as silica. Other elements, such as Na, are nonessential, but are deposited throughout the plant. Their concentration will depend entirely on extrinsic factors regulating their availability in the soil solution, i.e., moisture and soil content. Similarly, Cl content is determined less by the needs of the plant than by the availability in the soil solution; in addition to occurring naturally, Cl is present in excess as the anion complement in K fertilizer applications. An analysis was performed on existing data for switchgrass samples from ten different farms in the south-central portion of Iowa, with the goal of determining correlations between switchgrass elemental composition and geographical and seasonal changes so as to identify factors that influence the elemental composition of biomass. The most important factors in determining levels of various chemical compounds were found to be seasonal and geographical differences related to soil conditions. Combustion testing was performed to obtain deposits typical of boiler fouling and slagging conditions as well as fly ash. Analysis methods using computer-controlled scanning electron microscopy and chemical fractionation were applied to determine the composition and association of inorganic materials in the biomass samples. Modified sample preparation techniques and mineral quantification procedures using cluster analysis were developed to characterize the inorganic material in these samples. Each of the biomass types exhibited different inorganic associations in the fuel as well as in the deposits and fly ash. Morphological analyses of the wheat straw show elongated 10-30-{micro}m amorphous silica particles or phytoliths in the wheat straw structure. Alkali such as potassium, calcium, and sodium is organically bound and dispersed in the organic structure of the biomass materials. Combustion test results showed that the blends fed quite evenly, with good burnout. Significant slag deposit formation was observed for the 100% wheat straw, compared to bituminous and subbituminous coals burned under similar conditions. Although growing rapidly, the fouling deposits of the biomass and coal-biomass blends were significantly weaker than those of the coals. Fouling was only slightly worse for the 100% wheat straw fuel compared to the coals. The wheat straw ash was found to show the greatest similarity from the fuel to the ash analyzed. A high percentage of particles from both fuel and ash samples contained both Si and K. While Cl was a significant component in the fuel, very little was detected in the ash sample