Industry-relevant magnetron sputtering and cathodic arc ultra-high vacuum deposition system for in situ x-ray diffraction studies of thin film growth using high energy synchrotron radiation.

We present an industry-relevant, large-scale, ultra-high vacuum (UHV) magnetron sputtering and cathodic arc deposition system purposefully designed for time-resolved in situ thin film deposition/annealing studies using high-energy (>50 keV), high photon flux (>10(12) ph/s) synchrotron radiation. The high photon flux, combined with a fast-acquisition-time (<1 s) two-dimensional (2D) detector, permits time-resolved in situ structural analysis of thin film formation processes. The high-energy synchrotron-radiation based x-rays result in small scattering angles (<11°), allowing large areas of reciprocal space to be imaged with a 2D detector. The system has been designed for use on the 1-tonne, ultra-high load, high-resolution hexapod at the P07 High Energy Materials Science beamline at PETRA III at the Deutsches Elektronen-Synchrotron in Hamburg, Germany. The deposition system includes standard features of a typical UHV deposition system plus a range of special features suited for synchrotron radiation studies and industry-relevant processes. We openly encourage the materials research community to contact us for collaborative opportunities using this unique and versatile scientific instrument.

Biomarker selection for detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR.

Accurate identification of lymph node involvement is critical for successful treatment of patients with colorectal carcinoma (CRC). Real-time quantitative RT-PCR with a specific probe and RNA copy standard for biomarker mRNA has proven very powerful for detection of disseminated tumour cells. Which properties of biomarker mRNAs are important for identification of disseminated CRC cells? Seven biomarker candidates, CEA, CEACAM1-S/L, CEACAM6, CEACAM7-1/2, MUC2, MMP7 and CK20, were compared in a test-set of lymph nodes from 51 CRC patients (Dukes' A-D) and 10 controls. Normal colon epithelial cells, primary tumours, and different immune cells were also analysed. The biomarkers were ranked according to: (1) detection of haematoxylin/eosin positive nodes, (2) detection of Dukes' A and B patients, who developed metastases during a 54 months follow-up period and (3) identification of patients with Dukes' C and D tumours using the highest value of control nodes as cutoff. The following properties appear to be of importance; (a) no expression in immune cells, (b) relatively high and constant expression in tumour tissue irrespective of Dukes' stage and (c) no or weak downregulation in tumours compared to normal tissue. CEA fulfilled these criteria best, followed by CK20 and MUC2.

Histidine-rich domain of the knob protein of the human malaria parasite Plasmodium falciparum.

Membranes of erythrocytes infected with the human malaria parasite Plasmodium falciparum develop protrusions called knobs. These structures are essential for the survival of the parasite in the host, and their induction requires the synthesis of the knob protein by the parasite. We describe the isolation of a cDNA clone encoding the amino-terminal half of the knob protein. A cDNA library was constructed from RNA prepared from ring stages of a P. falciparum isolate that has retained its ability to induce knobs (knob+ phenotype). A synthetic oligonucleotide probe encoding polyhistidine was used to isolate the cDNA clone, which encodes the amino-terminal half of a polypeptide with all the known attributes of the knob protein. The gene is not transcribed in variants that do not synthesize the knob protein and thereby cannot induce knobs (knob- phenotype). The apparent lack of transcription in knob- variants is due to different mechanisms: although the gene is present in one knob- isolate, it has been deleted in a cloned knob- variant. The primary structure of the polypeptide deduced from a partial sequence of the cDNA is distinctly different from other malarial histidine-rich polypeptides. The amino-terminal sequence shows the characteristic features of a signal peptide. This is followed by a histidine-rich domain and a subsequent region which contains one histidine. Peptide map analysis of the knob protein is consistent with the structural features deduced from the sequence analysis of the cDNA.

Perceptibility of experimental and clinical lesions in the CT image with and without image processing.

The Ohio-Nuclear Delta 50 FS whole-body CT scanner and image filtering programs FI and SP:SMOOTH provided with the equipment were used to investigate how the perceptibility in the CT image of artificial lesions in a phantom and of liver lesions in patients was influenced by different reconstruction diameters, radiation doses and section thicknesses and by image filtering and different conditions of viewing. The perceptibility of both artificial and clinical lesions was considerably improved, especially in scans with a low radiation dose, when optimum viewing conditions and image filters were used.

Rapid zymogen activation and isolation of serine proteases from an individual mouse pancreas by affinity chromatography: genetical heterogeneity of chymotrypsins of Mus musculus.

A rapid method to prepare homogeneous fractions of the various chymotrypsins and trypsins from a single mouse pancreas (130-150 mg wet weight) is described. The method was applied to investigate intra-species variation on a molecular level using chymotrypsins as biochemical indicators. The conditions for optimal extraction of the zymogens in the homogenized pancreas have been studied. DNA had to be removed from the homogenate to obtain maximum chymotrypsin yields (approximately 1% of the wet weight of the pancreas). The activation was initiated by immobilized bovine trypsin that was removed by filtration. Then chymotrypsinogens in the homogenate were activated by mouse trypsin. After completed activation homogeneous chymotrypsins (one anionic and one cationic form) could be isolated in an one step analytical affinity chromatographic separation, using soybean trypsin inhibitor bound in Sepharose as a protease specific adsorbent. The end products were characterized by isoelectric focussing, amino acid composition, enzymatic parameters, molar extinction coefficient, and the number of polypeptide chains. Hereby, the existence of two chymotrypsinogen loci in the mouse genome could be demonstrated. Differences in structure and function between the corresponding enzymes from the two strains were found. This allelomorphism was verified in the crossing of the off-spring.