Synthesis Gas Biorefinery.

Synthesis gas or syngas is an intermediate, which can be produced by gasification from a variety of carbonaceous feedstocks including biomass. Carbon monoxide and hydrogen, the main constituents of syngas, can be subjected to a broad range of chemical and microbial synthesis processes, leading to gaseous and liquid hydrocarbon fuels as well as to platform and fine chemicals. Gasification of solid biomass differs from coal gasification by chemical composition, heating value, ash behavior, and other technical and biomass related issues. By thermochemical pre-treatment of lignocellulose as the most abundant form of biomass, for example, by torrefaction or fast pyrolysis, energy dense fuels for gasification can be obtained, which can be used in the different types of gasifiers available today. A number of pilot and demonstration plants exist, giving evidence of the broad technology portfolio developed so far. Therefore, a syngas biorefinery is highly flexible in regard to feedstock and product options. However, the technology is complex and does not result in competitive production costs today. Added value can be generated by suitable integration of thermochemical, biochemical, and chemical processes.

TRIM25 has a dual function in the p53/Mdm2 circuit.

P53 is an important tumor suppressor that, upon activation, induces growth arrest and cell death. Control of p53 is thus of prime importance for proliferating cells, but also for cancer therapy, where p53 activity contributes to the eradication of tumors. Mdm2 functionally inhibits p53 and targets the tumor suppressor protein for degradation. In a genetic screen, we identified TRIM25 as a novel regulator of p53 and Mdm2. TRIM25 increased p53 and Mdm2 abundance by inhibiting their ubiquitination and degradation in 26 S proteasomes. TRIM25 co-precipitated with p53 and Mdm2 and interfered with the association of p300 and Mdm2, a critical step for p53 polyubiquitination. Despite the increase in p53 levels, p53 activity was inhibited in the presence of TRIM25. Downregulation of TRIM25 resulted in an increased acetylation of p53 and p53-dependent cell death in HCT116 cells. Upon genotoxic insults, TRIM25 dampened the p53-dependent DNA damage response. The downregulation of TRIM25 furthermore resulted in massive apoptosis during early embryogenesis of medaka, which was rescued by the concomitant downregulation of p53, demonstrating the functional relevance of the regulation of p53 by TRIM25 in an organismal context.

Effects of VIE tagging and partial tissue sampling on the immune response of three-spined stickleback Gasterosteus aculeatus.

A 14 day experiment on effects of visible implant elastomer (VIE) tagging and spine-clipping of three-spined stickleback Gasterosteus aculeatus showed significant increases in immune response, particularly in the granulocyte:lymphocyte ratio, in both treatments and the sham control. A minimum two-week recovery after handling, anaesthesia, tagging and spine-clipping is recommended to minimize effect of manipulation on the immune system.

Disseminated Geotrichum candidum infection in a patient with relapsed acute myelogenous leukemia following allogeneic stem cell transplantation and review of the literature.

We describe a woman with relapsed acute myelogenous leukemia after allogeneic stem cell transplantation who developed disseminated Geotrichum candidum infection during chemotherapy-induced neutropenia. The isolate was susceptible to voriconazole, amphotericin B, and micafungin in vitro. We review the literature regarding invasive infections with G. candidum, which predominantly affect immunocompromised hosts, and discuss potential therapies for this rare pathogen.

Transposon-mediated enhancer trapping in medaka.

We tested the Sleeping Beauty transposable element for its ability to efficiently insert transgenes into the genome of medaka (Oryzias latipes), an important model system for vertebrate development. We show that the SB transposon efficiently mediates integration of a reporter gene into the fish germ line. In pilot experiments, we established 174 transgenic lines with a transgenesis efficiency of 32%. Transgenes are stably transmitted to, and expressed in, subsequent generations. Interestingly, the transgenic lines show novel expression patterns with temporal and spatial specificity at a rate of 12% (21/174), likely due to both, enhancing and silencing position effects. Furthermore, promoter-dependent GFP expression in injected fish embryos is tightly correlated with germ line transmission, facilitating easy selection of founder fish. Thus, the SB transposon/transposase system provides a highly efficient tool for transgenesis in general and for the generation of novel reporter gene expression patterns in particular.

Medaka eyeless is the key factor linking retinal determination and eye growth.

The complete absence of eyes in the medaka fish mutation eyeless is the result of defective optic vesicle evagination. We show that the eyeless mutation is caused by an intronic insertion in the Rx3 homeobox gene resulting in a transcriptional repression of the locus that is rescued by injection of plasmid DNA containing the wild-type locus. Functional analysis reveals that Six3- and Pax6- dependent retina determination does not require Rx3. However, gain- and loss-of-function phenotypes show that Rx3 is indispensable to initiate optic vesicle evagination and to control vesicle proliferation, by that regulating organ size. Thus, Rx3 acts at a key position coupling the determination with subsequent morphogenesis and differentiation of the developing eye.

A genetic screen for mutations affecting embryonic development in medaka fish (Oryzias latipes).

In a pilot screen, we assayed the efficiency of ethylnitrosourea (ENU) as a chemical mutagen to induce mutations that lead to early embryonic and larval lethal phenotypes in the Japanese medaka fish, Oryzias latipes. ENU acts as a very efficient mutagen inducing mutations at high rates in germ cells. Three repeated treatments of male fish in 3 mM ENU for 1 h results in locus specific mutation rates of 1.1-1.95 x10(-3). Mutagenized males were outcrossed to wild type females and the F1 offspring was used to establish F2 families. F2 siblings were intercrossed and the F3 progeny was scored 24, 48 and 72 h after fertilization for morphological alterations affecting eye development. The presented mutant phenotypes were identified using morphological criteria and occur during early developmental stages of medaka. They are stably inherited in a Mendelian fashion. The high efficiency of ENU to induce mutations in this pilot screen indicates that chemical mutagenesis and screening for morphologically visible phenotypes in medaka fish allows the genetic analysis of specific aspects of vertebrate development complementing the screens performed in other vertebrate model systems.

The conditional medaka mutation eyeless uncouples patterning and morphogenesis of the eye.

In early vertebrate eye development, the retinal anlage is specified in the anterior neuroectoderm. During neurulation, the optic vesicles evaginate from the lateral wall of the prosencephalon. Here we describe the temperature-sensitive mutation eyeless in the Japanese medakafish. Marker gene analysis indicates that, whereas, specification of two retinal primordia and proximodistal patterning takes place in the mutant embryo, optic vesicle evagination does not occur and subsequent differentiation of the retinal primordia is not observed. The mutation eyeless thus uncouples patterning and morphogenesis at early steps of retinal development. Temperature-shift experiments indicate a requirement for eyeless activity prior to optic vesicle evagination. Cell transplantation shows that eyeless acts cell autonomously.

An in situ hybridization screen for the rapid isolation of differentially expressed genes.

To rapidly isolate genes specifically expressed during medaka development we generated a cDNA library enriched for genes expressed in the head region of the developing embryo. Clones were spotted on filters automatically and preselected for abundantly expressed genes by hybridizing them with a probe derived from RNA of undifferentiated totipotent cells. Of the nonhybridizing clones 153 were chosen randomly and further analyzed by whole-mount in situ hybridization. There were 67 selected clones differentially expressed in the developing embryos, and 48 of these were expressed in the developing head. Differentially expressed genes were either of novel type or showed homology to known genes containing DNA binding motifs or to putative housekeeping genes.

Morphogenesis of the optic tectum in the medaka (Oryzias latipes): a morphological and molecular study, with special emphasis on cell proliferation.

We analyzed the medaka optic tectum (OT) morphogenesis by using 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry (with a new method we developed for pulse-labeling embryos) and in situ hybridization with three probes, two for recently cloned homeobox genes (Ol-Prx3 [Paired-Related-Homeobox3] and Ol-Gsh1 [Genetic-Screen-Homeobox1]) and one for Ol-tailless. The tectal anlage first appears as a sheet of proliferating cells expressing Ol-Gsh1 and Ol-tailless but not Ol-Prx3. Cells subsequently cease to proliferate in a superficial and rostral zone and begin to express Ol-Prx3. When tectal lamination begins, the proliferative zone (mpz) becomes restricted to a crescent at the OT medial, caudal, and lateral margin. This mpz functions throughout the fish's entire life. It produces cells that are added at the OT's edge as radial rows, spanning every layer of the OT. The cells of the mpz continue to express Ol-tailless in the adult, whereas Ol-Gsh1 expression is turned off. When superficial layers form, Ol-Prx3 expression becomes restricted to the underlying deep layer, where it persists in the adult. Ol-Prx3 seems to be a marker for the differentiation of a subset of deep cells and allows analysis of tectal lamination, whereas Ol-tailless and Ol-Gsh1 could be involved in the control of tectal cell proliferation. This study constitutes a first step toward molecular approach to OT development in anamniotes. We compare and discuss the expression patterns of the homologs of the genes studied, and more generally the morphogenetic patterns of the medaka tectum, with those encountered in other cortical structures and in other vertebrate groups.

Maximal inspiratory pressure following maximal exercise in trained and untrained subjects.

Previous investigators have demonstrated that 5-10 min of fatiguing exercise would lead to respiratory muscle fatigue in normal subjects. The purpose of this study was to determine if there was a differential inspiratory pressure response to maximal cycle ergometer exercise in trained and untrained subjects. Six highly trained cross country skiers and five untrained college students were studied prior to and 10, 60, and 120 s postexercise (incremental VO2max to exhaustion). On each occasion, maximal inspiratory pressure (MIP) was measured at the mouth from residual volume. Prior to exercise, the two groups had similar MIP values. After exercise, the sedentary subjects experienced significant decreases in MIP compared to the preexercise values. These decreases averaged 10%, 17%, and 13% at 10, 60, and 120 s postexercise, respectively. The skiers, on the other hand, showed no evidence of a decrease in MIP postexercise, with the postexercise values being slightly, but not significantly, higher than the preexercise values. From these results, we conclude that maximal exercise results in inspiratory muscle dysfunction in normal subjects but not in athletes training at or near elite levels. Thus, it appears that endurance exercise training induces an adaptive change in the inspiratory muscles that protects them from the acute loss of strength seen following exercise in normal subjects.

Caffeine, maximal power output and fatigue.

The purpose of this investigation was to determine the effects of caffeine ingestion on maximal power output and fatigue during short term, high intensity exercise. Nine adult males performed 15 s maximal exercise bouts 60 min after ingestion of caffeine (7 mg.kg-1) or placebo. Exercise bouts were carried out on a modified cycle ergometer which allowed power output to be computed for each one-half pedal stroke via microcomputer. Peak power output under caffeine conditions was not significantly different from that obtained following placebo ingestion. Similarly, time to peak power, total work, power fatigue index and power fatigue rate did not differ significantly between caffeine and placebo conditions. These results suggest that caffeine ingestion does not increase one's maximal ability to generate power. Further, caffeine does not alter the rate or magnitude of fatigue during high intensity, dynamic exercise.