Dynamic complex opto-magnetic holography.

Despite recent significant progress in real-time, large-area computer-generated holography, its memory requirements and computational loads will be hard to tackle for several decades to come with the current paradigm based on a priori calculations and bit-plane writing to a spatial light modulator. Here we experimentally demonstrate a holistic approach to serial computation and repeatable writing of computer-generated dynamic holograms without Fourier transform, using minimal amounts of computer memory. We use the ultrafast opto-magnetic recording of holographic patterns in a ferrimagnetic film with femtosecond laser pulses, driven by the on-the-fly hardware computation of a single holographic point. The intensity-threshold nature of the magnetic medium allows sub-diffraction-limited, point-by-point toggling of arbitrarily localized magnetic spots on the sample, according to the proposed circular detour-phase encoding, providing complex modulation and symmetrical suppression of upper diffractive orders and conjugated terms in holographically reconstructed 3-D images.

Expanded bed adsorption for recovery of renatured human recombinant interleukin 8 from Escherichia coli inclusion bodies.

Expanded bed adsorption is a chromatography technique based on stable fluidization, designed for large scale recovery of proteins directly from particulate-containing feedstocks. Described in this article is the use of expanded bed adsorption for recovery of renatured recombinant human interleukin 8 from Escherichia coli inclusion bodies, using a cation exchange adsorbent. The yield of interleukin 8 from the expanded bed was approximately 100% and the purification factor was approximately 4. Following each interleukin 8 purification a cleaning in place procedure was performed with a commercial cleaner. The results show that the adsorbent can be used at least 50 times without any significant loss of function.

Pilot scale recovery of recombinant annexin V from unclarified escherichia coli homogenate using expanded bed adsorption.

Expanded bed adsorption is a new downstream processing technique forcapture of proteins directly from unclarified feedstocks. Expanded bed adsorption reduces the number of operations in purification processes by combining clarification, concentration, and capture into one operation. It is based on stable fluidization and uses adsorbent particles with well-defined size and density distributions, together with columns designed to giveeven liquid flow distribution. The bed expands as the adsorbent particles are lifted by an upward liquid flow through the column. The behavior of the expanded bed is similar to a packed chromatography bed due to very little back-mixing of the adsorbent particles. The major benefit of using anexpanded bed is that adsorption can be carried out with unclarified feedstocks; there is no need for centrifugation or filtration to remove cells and debris. When the feedstock is applied, the target protein is captured by the adsorbent while cells and debris pass through the column unhindered. Washing is performed with the bed in an expanded mode, followed by elution of bound protein in a sedimented mode with downward flowDescribed in this article is the use of expanded bed adsorption for pilot scale recovery of recombinant human placental annexin V from an Escherichia coli ho mogenate. The description includes the whole procedure, from small-scale method optimization to pilot scale. The recovery of annexin V was approximately 95% at both lab scale and pilot scale. During the trials, it was discovered that the expanded bed was affected by the biomass content and viscosity of the homogenate. The upper limits for these parameters were therefore investigated further. For the E. coli used in the application described here, homogenates with biomass dry weightup to 5% and viscosities up to 10 mPa s (at a shear rate of 1 s(-1)) worked best. It was, however, feasible to use homogenates with dry weight up to 7-8% and viscosities up to 50 mPa s (1 s(-1)). (c) 1994 John Wiley & Sons, Inc.

[The triad obesity, hypertension and glycoregulation complaints in a suburban population of Tunisia's Sahel]. / La triade obésité, hypertension et troubles de la glycorégulation dans une population semi-urbaine du sahel Tunisien.

Obesity, hypertension and glucose intolerance are often related, the relationship between hypertension and glycemia is well documented and may be explained through hyperinsulinemia. Most of the studies were conducted in western societies. The simultaneous study of this triad in a random sample (n = 555) of a suburban adult community of Tunisian's Sahel, showed an association between hypertension and level of overweight which was also related to diabetes prevalence after matching for age and sex. However serum glucose was related to both systolic and diastolic blood pressure only for non obese subjects, when levels of overweight are stratified. The association between diabetes and hypertension therefore involves different mechanisms for obese and non obese subjects.

Cloning and characterization of the t(15;17) translocation breakpoint region in acute promyelocytic leukemia.

A reciprocal chromosomal translocation, t(15;17)(q22;q11.2-12), is characteristic of acute promyelocytic leukemia (APL) of French-American-British (FAB) subtype M3, and is not associated with any other human malignancy. The non-random pattern of the APL translocations suggests that specific genes on chromosomes 15 and 17 are somehow altered or deregulated as a consequence of the rearrangement. Translocation breakpoints in APL patients provide physical landmarks that suggest an approach to isolating the APL gene(s). Genetic and physical maps constructed for the APL breakpoint region on chromosome 17 have indicated that two fully-linked DNA markers, defining loci for THRA1 and D17S80, map to opposite sides of an APL breakpoint yet reside on a common 350-kb Clal fragment. Cosmid-walking experiments to clone this APL breakpoint have revealed a 38-kilobase deletion on chromosome 17. Studies in additional APL patients have shown that the breakpoint region on chromosome 17 spans at least 80 kilobases.

Fine structure DNA mapping studies of the chromosomal region harboring the genetic defect in neurofibromatosis type I.

To better map the location of the von Recklinghausen neurofibromatosis (NF1) gene, we have characterized a somatic cell hybrid designated 7AE-11. This microcell-mediated, chromosome-transfer construct harbors a centromeric segment and a neo-marked segment from the distal long arm of human chromosome 17. We have identified 269 cosmid clones with human sequences from a 7AE-11 library and, using a panel of somatic cell hybrids with a total of six chromosome 17q breakpoints, have mapped 240 of these clones on chromosome 17q. The panel included a hybrid (NF13) carrying a der(22) chromosome that was isolated from an NF1 patient with a balanced translocation, t(17;22) (q11.2;q11.2). Fifty-three of the cosmids map into a region spanning the NF13 breakpoint, as defined by the two closest flanking breakpoints (17q11.2 and 17q11.2-q12). RFLP clones from a subset of these cosmids have been mapped by linkage analysis in normal reference families, to localize the NF1 gene more precisely and to enhance the potential for genetic diagnosis of this disorder. The cosmids in the NF1 region will be an important resource for testing DNA blots of large-fragment restriction-enzyme digests from NF1 patient cell lines, to detect rearrangements in patients' DNA and to identify the 17;22 NF1 translocation breakpoint.

Structure of the IgG-binding regions of streptococcal protein G.

The gene encoding the IgG-binding protein G from Streptococcus G148 was isolated by molecular cloning. A subclone containing a 1.5-kb insert gave a functional product in Escherichia coli. Protein analysis of affinity-purified polypeptides revealed two gene products, both smaller than protein G spontaneously released from streptococci, but with identical IgG-binding properties. The complete nucleotide sequence of the insert revealed a repeated structure probably evolved through duplications of fragments of different sizes. The deduced amino acid sequence revealed an open reading frame extending throughout the insert, terminating in a TAA stop codon. Analysis of the two gene products by N-terminal amino acid determination suggests that two different TTG codons are recognized in E. coli for initiation of translation to yield the two products. Based on these results several truncated gene constructions were expressed and analysed. The results suggest that the C-terminal part of streptococcal protein G consists of three IgG-binding domains followed by a region which anchors the protein to the cell surface. Structural and functional comparisons with streptococcal M protein and staphylococcal protein A have been made.

Monitoring of protein product formation during fermentation with fast protein liquid chromatography.

A method is described using fast protein liquid chromatography (FPLC) for the monitoring of protein formation during fermentation. The procedure consists of centrifugation to recover the cells, sonication of the cells, centrifugation to remove cell debris, and analysis of supernatant on a column of Mono Q (a strong anion exchanger). Analysis of peak areas provides quantitative determination of product concentration. Maintenance and life of the Mono Q column is discussed. We find that FPLC is a convenient method for measuring products in cell homogenates because it gives rapid, highly resolved separations.