Risk assessment for patients with chronic myeloid leukaemia before allogeneic blood or marrow transplantation. Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation

BACKGROUND: Transplantation of blood or bone-marrow stem cells is the treatment of choice for selected patients with chronic myeloid leukaemia (CML). Transplantation is used with increasing frequency and success, but remains associated with substantial risks of morbidity and mortality. Other treatments with satisfactory short-term outcome are available. For appropriate counselling of patients, a rapid and simple way to assess risk is needed. METHODS: Data from 3142 patients (1873 [60%] male, 1269 [40%] female; mean age 34 years, range <1-60 years) treated with allogeneic blood or marrow transplants for CML between 1989 and 1997, reported to the European Group for Blood and Marrow Transplantation (EBMT), were used to develop and test a simple risk score based on previously reported major pretransplant risk factors: histocompatibility, stage of disease at time of transplantation, age and sex of donor and recipient, and time from diagnosis to transplantation. We analysed probabilities of survival, leukaemia-free survival, transplant-related mortality, and relapse incidence with respect to these risk factors. FINDINGS: At the time of analysis, 1922 (61%) of the 3142 patients were alive-1567 (65%) of those with HLA-identical sibling donors and 417 (57%) of those with unrelated donors. 1682 (54%) were alive without relapse. 1220 (39%) patients had died, 1013 (83%) of transplant-related causes, 207 (17%) of relapse. 447 (14%) patients had relapsed. The final scoring system was highly predictive for leukaemia-free survival, survival and transplant-related mortality. Survival at 5 years was 72%, 70%, 62%, 48%, 40%, 18%, and 22% for patients with scores 0, 1, 2, 3, 4, 5, and 6, respectively. Risk of transplant-related mortality was 20%, 23%, 31%, 46%, 51%, 71%, and 73%. Data showed the same trends for HLA-identical sibling transplants and unrelated transplants for transplants done in 1989-93 and 1994-96. INTERPRETATION: Pretransplant risk factors are cumulative for individual patients with CML having blood or marrow transplantation. A simple system based on five main factors gives adequate risk assessment for counselling of patients and taking decisions.     

Nonradioactive Northern blotting with biotinylated and digoxigenin-labeled RNA probes

The application of nonradioactive RNA probes for Northern blotting offers the advantage of a rapid turn-around time for results without the loss of sensitivity for target mRNA detection. However, a problem that has impeded the widespread use of nonradioactive RNA probes for use in Northern blotting is the difficulty in stripping these probes from nylon membranes after hybridization. In this report we describe two protocols for stripping digoxigenin (Dig)-labeled RNA probes from nylon membranes. One protocol utilizes a phosphate-buffered formamide stripping solution to remove nonchemically modified (regular) RNA probes while the other method utilizes strippable probes that were produced with a chemically modified nucleotide (CTP) and removed by a specific stripping solution. This latter method was developed by Ambion Inc. and is called Strip-EZ. We also describe a protocol for the detection of two separate rat mRNAs using both biotin and digoxigenin-labeled RNA probes that does not require stripping the membrane after hybridization. Finally, we describe the use of another new labeling technology, called Chem-Link, that quickly and conveniently labels RNA for use in Northern blotting.     

A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity

A novel chitinase gene of tobacco was isolated and characterized by DNA sequence analysis of a genomic clone and a cDNA clone. Comparative sequence analysis of both clones showed an identity of 94%. The proteins encoded by these sequences do not correspond to any of the previously characterized plant chitinases of classes I-IV and are designated as class V chitinases. Comparison of the chitinase class V peptide sequence with sequences in the Swiss Protein databank revealed significant sequence similarity with bacterial exo-chitinases from Bacillus circulans, Serratia marcescens and Streptomyces plicatus. It was demonstrated that class V chitinase gene expression is induced after treatment of tobacco with different forms of stress, like TMV-infection, ethylene treatment, wounding or ultraviolet irradiation. Two related chitinase class V proteins of 41 and 43 kDa were purified from Samsun NN tobacco leaves inoculated with tobacco mosaic virus. The proteins were purified by Chelating Superose chromatography and gel filtration. In vitro assays demonstrated that class V chitinases have endo-chitinase activity and exhibit antifungal activity toward Trichoderma viride and Alternaria radicina. In addition, it was shown that class V chitinase acts synergistically with tobacco class I beta-1,3-glucanase against Fusarium solani germlings.     

Complexities in STEM analyses of polymer blend thin films

Polystyrene (PS)-polyether sulphone (PES) polymer blend thin films were prepared for examination in a scanning transmission electron microscope. The microstructures observed in 75 wt % PS-25 wt % PES films consisted of spherical inclusions, ranging from 0.2 to 1.2m in diameter. X-ray spectrometric analysis in the microscope revealed that the inclusions were PES-rich, while the matrix contained only PS. Attention in this paper is paid to the contrast in the annular dark-field detector (ADF) images from these thin films. This image contrast has a complicated dependence on both the angular range subtended by the dark-field detector and mass-thickness variations within the films. On microscopes with appropriate lens controls which permit the acceptance angle of the ADF detector to be varied, it becomes possible actually to reverse the contrast between the two phases.     

Microdeformation in partially compatible blends of poly(styrene-acrylonitrile) and polycarbonate

Dilute 3-component 1-phase solutions in methylene chloride of poly(styrene-acrylonitrile) PSAN and polycarbonate PC are used to cast phase separated thin films. Films of pure PSAN, pure PC and five intermediate compositions are examined. The films are bonded to copper grids and strained at a constant rate of 4.1 × 10^–6sec^–1. The median tensile strain _v for void formation is determined using an optical microscope and the regions surrounding the voids are examined by TEM. At room temperature and slow strain rates both PSAN and PC plastically deform by shear yielding. For pure PSAN _v was found to be 0.13 whereas for PC _v exceeds 0.23. The addition of the more ductile polymer PC to PSAN at weight fractionsx forx 0.4 decreases _v. In this case voids form in crazes at the boundaries between the PC-rich inclusion and the PSAN-rich matrix. When the PC content is increased tox = 0.8, _v approaches 0.23. The effect of physical ageing (annealing belowT _g the glass transition temperature) on the mode of plastic deformation was also examined over the same compositional range. Physical ageing was found to suppress shear deformation and favour crazing in PSAN and PSAN-rich phases. Because crazes are more susceptible to breakdown than DZ's (shear deformation zones), physical ageing results in a marked decrease in _v. The breakdown statistics of these phase separated partially compatible blends was found to follow a Weibull distribution in strain from which two parameters may be extracted: the Weibull modulus and _w the Weibull scale parameter. is a measure of the breadth of distribution of void initiation and _w is a measure of the median strain to void formation in the films. The behaviour of _w was found to approximately mirror _v. The Weibull modulus appears to be primarily controlled by the matrix phase.     

Cold drawing of polyethylene terephthalate/polyethylene films

Films made from a 77/23 weight percent polyethylene terephthalate/polyethylene blend were stretched using an Instron apparatus. The structures of the samples were examined by methods which included ion beam etching and electron microscopy. The drawn samples exhibited a metallically lustrous and highly reflective surface up to a draw temperature, T_R, of 70°C and a draw ratio of about 5. Under certain conditions the change in structure caused by the stretching resulted in the formation of a highly uniform fibril-void structure. The voids are up to 3 μm in width and 160 μm in length and are formed primarily as a result of aggregation and then separation of the microfibrils of the individual polymer components.     

Enzymatic esterification of glycerol II. Lipase-catalyzed synthesis of regioisomerically pure 1(3)-rac-monoacylglycerols

Regioisomerically pure 1(3)-rac-monoacylglycerols are conveniently prepared in high yields (>75%) and in multigram quantities by enzymatic esterification of glycerol in the presence of various lipases(Chromobacterium viscosum, Rhizopus delemar, Rhizomucor miehei) with a variety of different acyl donors, such as free fatty acids, fatty acid alkyl esters, vinyl esters and triacylglycerols, as well as natural fats and oils. All reactions are carried out in aprotic organic solvents with low water content, namelyn-hexane, diethyl ether, tBuOMe or mixtures of these solvents. Essential for the success of these transformations were the following two factors. First, the creation of an artificial interphase between the solvent-immiscible hydrophilic glycerol and the hydrophobic reaction medium by its adsorption onto a solid support. Second, a facile system for the separation of the desired monoacylglycerol from the reaction mixture, coupled with the continuous recycling of acyl donor and undesirable by-products.     

Enzymatic esterification of glycerol I. Lipase-catalyzed synthesis of regioisomerically pure 1,3-sn-diacylglycerols

Regioisomerically pure 1,3-sn-diacylglycerols are conveniently prepared in high yields (>80%) and in large quantities by enzymatic esterification of glycerol in the presence of various 1,3-selective lipases(Chromobacterium viscosum, Rhizopus delemar, Rhizomucor miehei) and a variety of different acyl donors like free fatty acids, fatty acid alkyl esters and vinyl esters. All reactions are carried out in aprotic organic solvents of low water content, namelyn-hexane, diethyl ether or tBuOMe. The creation of an artificial interphase between the solvent-immiscible hydrophilic glycerol and the hydrophobic reaction media by the adsorption of glycerol onto a solid support prior to use was essential for the success of these transformations. The effects of reaction conditions and the regioselectivities of the lipases on the product yields are described in detail.     

Light-sensitive elastomers modified by 4-(N-dimethylmaleimido)-benzenesulphenyl chloride

New light-sensitive polymers were prepared by modification of four different elastomers with 4-(N-dimethylmaleimido)-benzenesulphenyl chloride (DMI-BSCI). Polydienes modified by DMI-BSCI form block copolymers. These polymers show a strong increase of the glass transition temperature T_g whereby the T_g increase of modified poly(octenamer) is much smaller. Higher modified polydienes have two T_g. Modified poly(octenamers) show a melting temperature dependence T_m on the content of trans double bonds which is characteristic of statistical copolymers. The ratio of rate constants with which trans- and cis-structures in poly(octenamer) are occupied by DMI-BSCI, is

$\text{k}{}_{\mathrm{trans}}\text{k}{}_{\mathrm{cis}}\text{= 0.34}$

     

机械总院研发立式冷摆动辗压机填补国内空白

机械科学研究总院北京机电研究所承担“2000KN立式冷摆动辗压机开发研究”项目通过验收,研发的2000kN矿式冷摆动辗压机具有自主知识产权,填补了国内空白,达到国际先进水平,人选“2013度中关村科技园区首台（套）重大技术装备试验、示范项目。