Peroxisome-deficient mutants of Hansenula polymorpha

As a first step in a genetic approach towards understanding peroxisome biogenesis and function, we have sought to isolate mutants of the methylotrophic yeast Hansenula polymorpha which are deficient in peroxisomes. A collection of 260 methanol-utilization-defective strains was isolated and screened for the ability to utilize a second compound, ethanol, the metabolism of which involves peroxisomes. Electron microscopical investigations of ultrathin sections of selected pleiotropic mutants revealed two strains which were completely devoid of peroxisomes. In both, different peroxisomal matrix enzymes were active but located in the cytosol; these included catalase, alcohol oxidase, malate synthase and isocitrate lyase. Subsequent backcrossing experiments revealed that for all crosses involving both strains, the methanol- and ethanol utilizing-deficient phenotypes segregated independently of each other, indicating that different gene mutations were responsible for these phenotypes. The phenotype of the backcrossed peroxisome-deficient derivates was identical: defective in the ability to utilize methanol but capable of growth on other carbon sources, including ethanol. The mutations complemented and therefore were recessive mutations in different genes.     

Approximation of theoretical energy-saving potentials for the petrochemical industry using energy balances for 68 key processes

We prepared energy and carbon balances for 68 petrochemical processes in the petrochemical industry for Western Europe, the Netherlands and the world. We analysed the process energy use in relation to the heat effects of the chemical reactions and quantified in this way the sum of all energy inputs into the processes that do not end up in the useful products of the process, but are lost as waste heat to the environment. We showed that both process energy use and heat effects of reaction contribute significantly to the overall energy loss of the processes studied and recommend addressing reaction effects explicitly in energy-efficiency studies. We estimated the energy loss in Western Europe in the year 2000 at 1620 PJ of final energy and 1936 PJ of primary energy, resulting in a total of 127 Mt CO₂. The losses identified can be regarded as good approximations of the theoretical energy-saving potentials of the processes analysed. The processes with large energy losses in relative (per tonne of product) and absolute (in PJ per year) terms are recommended for more detailed analysis taking into account further thermodynamic, economic, and practical considerations to identify technical and economic energy-saving potentials.     

Development of hardware around a silicon microstrip detector with 512 elements

The silicon chip is mounted on a ceramic substrate and connected to the preamplifiers via kapton foils. A test bench is constructed, which enables measurement of reverse current and capacitance of all strips simultaneously.     

Functional mimicry of a discontinuous antigenic site by a designed synthetic peptide

Functional reproduction of the discontinuous antigenic site D of foot-and-mouth disease virus (FMDV) has been achieved by means of synthetic peptide constructions that integrate each of the three protein loops that define the antigenic site into a single molecule. The site D mimics were designed on the basis of the X-ray structure of FMDV type C-S8c1 with the aid of molecular dynamics, so that the five residues assumed to be involved in antigenic recognition are located on the same face of the molecule, exposed to solvent and defining a set of native-like distances and angles. The designed site D mimics are disulfide-linked heterodimers that consist of a larger unit containing VP2(71-84), followed by a polyproline module and by VP3(52-62), and a smaller unit corresponding to VP1(188-194) (VP=viral protein). Guinea pig antisera to the peptides recognized the viral particle and competed with site D-specific monoclonal antibodies, while inoculation with a simple (not covalently joined to one another) admixture of the three VP1-VP3 sequences yielded no detectable virus-specific serum conversion. Similar results have been reproduced in two bovines. Antisera to the peptides also moderately neutralize FMDV in cell cultures and partially protect guinea pigs against challenge with the virus. These results demonstrate functional mimicry of the discontinuous site D by the peptides, which are therefore obvious candidates for a multicomponent, peptide-based vaccine against FMDV.     

Coumarins inPrunus mahaleb and its herbivore,the small ermine mothYponomeuta mahalebellus

Larvae of the small ermine mothYponomeuta mahalebellus were reared on foliage ofPrunus mahaleb, a plant known to contain coumarins. Thin-layer chromatography and gas chromatography showed that coumarin, umbelliferone, and herniarin were present in leaves ofP. mahaleb and in pupae and adults ofY. mahalebellus. Overall concentrations of simple coumarins in the plant and insect were, respectively, 0.54% and 0.003–0.004% (dry weight). The possible role of coumarins in the chemical defense of both the plant and insect is discussed.     

Anatomical-based FDG-PET reconstruction for the detection of hypo-metabolic regions in epilepsy

Positron emission tomography (PET) of the cerebral glucose metabolism has shown to be useful in the presurgical evaluation of patients with epilepsy. Between seizures, PET images using fluorodeoxyglucose (FDG) show a decreased glucose metabolism in areas of the gray matter (GM) tissue that are associated with the epileptogenic region. However, detection of subtle hypo-metabolic regions is limited by noise in the projection data and the relatively small thickness of the GM tissue compared to the spatial resolution of the PET system. Therefore, we present an iterative maximum-a-posteriori based reconstruction algorithm, dedicated to the detection of hypo-metabolic regions in FDG-PET images of the brain of epilepsy patients. Anatomical information, derived from magnetic resonance imaging data, and pathophysiological knowledge was included in the reconstruction algorithm. Two Monte Carlo based brain software phantom experiments were used to examine the performance of the algorithm. In the first experiment, we used perfect, and in the second, imperfect anatomical knowledge during the reconstruction process. In both experiments, we measured signal-to-noise ratio (SNR), root mean squared (rms) bias and rms standard deviation. For both experiments, bias was reduced at matched noise levels, when compared to post-smoothed maximum-likelihood expectation-maximization (ML-EM) and maximum a posteriori reconstruction without anatomical priors. The SNR was similar to that of ML-EM with optimal post-smoothing, although the parameters of the prior distributions were not optimized. We can conclude that the use of anatomical information combined with prior information about the underlying pathology is very promising for the detection of subtle hypo-metabolic regions in the brain of patients with epilepsy.