Morphology and agglomeration control of LiMnPO4 micro- and nanocrystals.

Microwave-assisted hydrothermal synthesis was used to grow LiMnPO4 micro- and nanocrystals from acetate precursors. By appropriate adjustment of the precursor concentration and the pH-value of the reactant, the product composition and purity along with the crystal size can be manipulated, resulting in particle-dimensions from around 10 µm down to a few 100 nm. Prisms and plates with hexagonal basal faces as well as cuboid and rod-like particles were produced. The effects on the crystal morphology as well as on the materials texture and agglomeration tendency are discussed and a comprehensive agglomeration phase diagram is constructed.

Combined use of fluorescent dyes and flow cytometry to quantify the physiological state of Pichia pastoris during the production of heterologous proteins in high-cell-density fed-batch cultures.

Matching both the construction of a recombinant strain and the process design with the characteristics of the target protein has the potential to significantly enhance bioprocess performance, robustness, and reproducibility. The factors affecting the physiological state of recombinant Pichia pastoris Mut(+) (methanol utilization-positive) strains and their cell membranes were quantified at the individual cell level using a combination of staining with fluorescent dyes and flow cytometric enumeration. Cell vitalities were found to range from 5 to 95% under various process conditions in high-cell-density fed-batch cultures, with strains producing either porcine trypsinogen or horseradish peroxidase extracellularly. Impaired cell vitality was observed to be the combined effect of production of recombinant protein, low pH, and high cell density. Vitality improved when any one of these stress factors was excluded. At a pH value of 4, which is commonly applied to counter proteolysis, recombinant strains exhibited severe physiological stress, whereas strains without heterologous genes were not affected. Physiologically compromised cells were also found to be increasingly sensitive to methanol when it accumulated in the culture broth. The magnitude of the response varied when different reporters were combined with either the native AOX1 promoter or its d6* variant, which differ in both strength and regulation. Finally, the quantitative assessment of the physiology of individual cells enables the implementation of innovative concepts in bioprocess development. Such concepts are in contrast to the frequently used paradigm, which always assumes a uniform cell population, because differentiation between the individual cells is not possible with methods commonly used.

Swiss Industrial Biocatalysis Consortium (SIBC).

Taking up the common challenges in biocatalysis, a group of industrialists decided to react with a bottom-up solution, and created the Swiss Industrial Biocatalysis Consortium (SIBC). The Swiss Industrial Biocatalysis Consortium is a pre-competitive working group to better implement and utilize existing know-how and resources in biocatalysis, and to influence and shape the economic and educational political environment. Recent examples of activities are outlined.

Arterial calcification in patients with chronic kidney disease.

OBJECTIVE: In patients with chronic kidney disease (CKD), aortic calcification is more frequent and severe and it is also predictive of adverse cardiovascular outcome. The aim of the present study was to characterize aortic calcification in renal compared with non-renal patients. METHODS: Aortas of 31 patients with advanced CKD and of 31 age-and gender-matched controls were obtained at autopsy. Calcium and phosphorus content in the aorta was quantitated using x-ray analysis. The expression of calcification-promoting and calcification-inhibiting proteins was assessed using immunohistochemistry. RESULTS: The calcium and phosphorus content of the aorta was higher in CKD patients than in controls. Even in non-calcified aortic specimens of CKD, staining for Msx-2, BMP-2, bone sialo-protein, TNF-alpha and nitrotyrosine was significantly more marked compared to controls. The same proteins were immunodetected in calcified aortic specimens of both CKD and controls. In contrast, staining for transglutaminase-2 and Fetuin A was significantly reduced in CKD. Higher expression of cbfa-1 and Pit-1 was observed in all calcified aortas with no difference between CKD and controls. The expression of TNF-alpha, phospho-p38 and Msx-2 was correlated to the intensity of upregulation of BMP-2 and osteoblastic transdifferentiation by VSMC even in non-calcified areas of the aortas of CKD. CONCLUSION: The expression of markers characteristic for calcification is not different in calcified aorta of CKD patients compared to controls, but in CKD patients, evidence of inflammation, transformation to an osteoblastic phenotype and reduced expression of transglutaminase are also found even in non-calcified aorta.

Calcification of coronary intima and media: immunohistochemistry, backscatter imaging, and x-ray analysis in renal and nonrenal patients.

Coronary calcification is a potent predictor of cardiac events. In patients with chronic renal disease, both prevalence and intensity of coronary calcification are increased. It has remained uncertain whether it is the intima of the coronaries or the media that is calcified and whether the morphologic details of calcified plaques differ between renal and nonrenal patients. Autopsy samples of coronaries were obtained from standard sites in 23 renal and 23 age- and gender-matched nonuremic patients. Specimens were examined using light and electron microscopy, immunohistochemistry, backscatter imaging, and x-ray analysis. In coronaries, calcified plaques occupied a similar proportion of the intima area in renal versus nonrenal patients (17.3 +/- 11.9 versus 18.1 +/- 11.9%) but occupied a significantly higher proportion of the media (16.6 +/- 10.6 versus 3.8 +/- 2.31%). Expression of the proteins osteocalcin, C-reactive protein, TGF-beta, and collagen IV was significantly more intensive around coronary plaques of renal compared with nonrenal patients. The non-plaque-bearing intima of renal patients showed minimal staining for fetuin, but fetuin staining was seen surrounding calcified plaques. In addition, more pronounced deposition of C5b-9 was found around coronary plaques of renal patients, and glycophorin deposition pointed to more past intraplaque hemorrhage in renal patients. Calcification by electron backscatter analysis is more intense in the coronary media, but not if the intima is more intense in renal compared with nonrenal patients. A more marked inflammatory response in renal patients is suggested by more frequent presence and greater intensity of markers of inflammation.