A description is given of a fully automatic program, written in ALGOL 60, that finds the constants of the reciprocal lattice from powder data. The progress of the program is illustrated with the (nearly) complete computer output for one selected case. Planes through the origin of the reciprocal lattice (zones) are found first. After evaluating these, the program selects pairs of zones with a common row in order to find reciprocal lattices, which are then reduced in a simple way. Each solution is compared with the experimental data and a figure of merit is calculated. The program is most suited for compounds of orthorhombic or lower symmetry.
A method is presented of determining powder data from photographs. With a microphotometer the density is measured in steps of 0.01 ° 20. These data are used by a computer program. With this program the background and the noise level are determined first. After the detection of the diffraction peaks, the peak positions and intensities are refined, a suitable model for the peak profile being used. A procedure is described for scaling of several photographs. The final results are illustrated with a comparison of 20 values determined according to this method and the calculated values of the lattice refined with these data. The resulting data prove to be equivalent to the data obtained by conventional means. The computer program is written in FORTRAN IV.
A method described to purify pluripotent hemopoietic stem cells ( PHSC ) from adult mouse bone marrow. The method consists of three separation steps. First, bone marrow cells are centrifuged in a discontinuous metrizamide gradient and simultaneously labeled with wheat germ agglutinin-fluorescein isothiocyanate (WGA-FITC). Second, the low density cells are analyzed by a fluorescence-activated cell sorter (FACS) and the WGA-positive cells with medium forward and low perpendicular light scatter intensities are sorted. The WGA-FITC is removed from the cells by incubation with N-acetyl-D-glucosamine. Finally, the sorted cells are incubated with anti-H-2K-biotin and avidin-FITC and sorted a second time to enrich cells with high H-2K density. The sorted cells gave rise to 2 spleen colonies per 100 injected cells at 8 d and 6.6 colonies per 100 cells at 12 d after transplantation into lethally irradiated syngeneic recipients. The average enrichment factor for day 12 CFU-S (colony-forming unit/spleen) was 135 (range, 90--230; n = 15) and was similar to that for the cell type that provides radioprotection (180 +/- 70), indicating that these functional properties were copurified. Indirect evidence suggests that the spleen-seeding efficiency (f factor) of these cells is 0.10 and, therefore, the average purity of the sorted PHSC was 65% (range in 15 experiments, 35--110%). The sorted cells were all in the G1 or G0 phase of the cell cycle. They appeared to be undifferentiated blasts by morphological criteria. Electron microscopy revealed that the sorted cells consisted primarily of two cell types, possibly representing G0 and G1 cells. The FACS was used to deposit single selected cells into individual microwells of Terasaki trays. 32% of the sorted cells could be induced to form myeloid progeny in vitro. This procedure should be useful for direct studies on the regulation of hemopoietic cell differentiation.
We have synthesized three peptides from the mdm-2 binding domain of human p53, residues 12-26 (PPLSQETFSDLWKLL), residues 12-20, and 17-26. To enable transport of the peptides across the cell membrane and at the same time to maximize the active mdm-2 binding ␣-helical conformation for these peptides, each was attached at its carboxyl terminus to the penetratin sequence, KKWKMRRNQF-WVKVQRG, that contains many positively charged residues that stabilize an ␣-helix when present on its carboxyl terminal end. All three peptides were cytotoxic to human cancer cells in culture, whereas a control, unrelated peptide attached to the same penetratin sequence had no effect on these cell lines. The same three cytotoxic peptides had no effect on the growth of normal cells, including human cord blood-derived stem cells. These peptides were as effective in causing cell death in p53-null cancer cells as in those having mutant or normal p53. Peptide-induced cell death is not accompanied by expression of apoptosis-associated proteins such as Bax and waf p21 . Based on these findings, we conclude that the antiproliferative effects of these p53-derived peptides are not completely dependent on p53 activity and may prove useful as general anticancer agents.
It is well established that the osteoclast is formed by fusion of post-mitotic, mononuclear precursors derived from circulating progenitor cells. However, the precise haematopoietic origin of the osteoclast is unknown. We have investigated this here by fractionating mouse bone marrow and isolating haematopoietic stem cells using a three-step method combining equilibrium density centrifugation and two fluorescence-activated cell sortings (FACS), and have tested the ability of each bone marrow fraction, including highly purified haematopoietic stem cells, to generate osteoclasts during co-culture with preosteoclast-free embryonic long bones. The osteoclast-forming capacity was found to increase with increasing stem cell purity. On the other hand, the culture time needed for osteoclast formation also increased with purification, suggesting the presence of progressively more immature progenitor cells. The pluripotent haematopoietic stem cell fractions with the highest purity needed preincubation with a stem cell-activating factor (interleukin-3) to activate the predominantly quiescent stem cells in vitro.
The elements of the unit cell are expressed in the coefficients A⋯F of the quadratic form 1/d2 = Q(h,k,l), and a table of derivatives δa/δA⋯δ cos γ/δF is given. The elements of Delaunay's foursider are calculated in a simple way from A⋯F.
A novel approach to the determination of crystallite size and lattice strain by means of Total Pattern Analysis is described. Parameters to define the position, magnitude, breadth and shape of individual peaks are obtained by an adaptation of the pattern fitting program of Sonneveld and Visser (J. Appl. Cryst. 8, 1-7, 1975). A rapid assessment of the nature of the specimen broadening is given by a Williamson-Hall Plot. This leads to a more detailed study of line breadths by, for example, Voigt analysis applied to several orders of reflections or to single lines. Preliminary results are given for the application of this procedure to 'size only' and 'size-strain' samples of ZnO.
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