Identification and characterization of human metabolites of CAI [5-amino-1-1(4''-chlorobenzoyl-3,5-dichlorobenzyl)-1,2,3-triazole- 4-carboxamide)

BACKGROUND: Bronchoscopic bronchoalveolar lavage in children to investigate bronchial disorders such as asthma has both ethical and procedural difficulties. OBJECTIVE: The aim of this study was to establish a standardized non-bronchoscopic method to perform bronchoalveolar lavage in children attending for elective surgery to obtain normal cellular data. METHODS: Bronchoalveolar lavage was performed on normal children (n = 55) by infusing saline (20 mL) through an 8 FG suction catheter passed after endotracheal intubation. Oxygen saturation, heart and respiratory rate were monitored during the bronchoalveolar lavage procedure. Cellular analysis and total protein estimation of the lavage fluid were performed. Epithelial lining fluid volume was calculated (n = 15) using the urea dilution method. RESULTS: The procedure was well tolerated by all children. Total cell count and differential cell count for children (macrophages 70.8 +/- 2.3%, lymphocytes 3.8 +/- 0.6%, neutrophils 5.7 +/- 1.0%, eosinophils 0.14 +/- 0.03%, epithelial cells 19.6 +/- 2.1%, mast cells 0.21 +/- 0.02%) were similar to those reported for adults. Age and sex comparisons revealed no differences between groups. The mean total protein recovered in the cell free supernatant was 49.72 +/- 4.29 mg/L and epithelial lining fluid volume was 0.82 +/- 0.11% of return lavageate. CONCLUSION: This method allows bronchoalveolar lavage to be performed safely and quickly on children attending for routine elective surgery. Using this method and taking the 'window of opportunity' of elective surgery, the presence or absence of airway inflammation could be studied in children with various patterns of asthma during relatively asymptomatic periods.     

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.     

Unusual W-shaped Galvanic Cell Model of ISCC of α-brass in Neutral Mattsson''''s Solution

Intergranular stress corrosion cracking(ISCC) of α-brass in neutral Mattsson's solutionwas found to be controlled by an unusual"W"-shaped galvanic cell whose cathode is thegrain boundary oxide film (G.B.0. film) andsurface film and the anode is fresh metal atthe cracked tip on both sides of the G.B.0.film. Redox reactions involved in the cellhave been proposed here. According to thismdel, initidtion of ISCC is caused by therupturing of surface film along grain boundaries,thus forming a galvanic cell. Propagation ofISCC resulted from alternate advances of G.B.0.film and dissolution on both sides of G.B.0.film caused by the effect of electrochemicalreaction. This work developed an effective approachto investigate the embrittlement process atthe tip of the crack, by increasing the lengthof the embrittlement region through constantstrain test and distinguishing the morphologyand the nature of the corrosion products byoptical microscopy and scanning electronmicroscopy (SEH).     

Loss of entanglement density during crazing

The formation of fibril surface area during craze growth requires a loss of entangled strand density in the fibrils themselves. To demonstrate the decrease in entangled chain density, thin films of polystyrene are bonded to soft copper grids and strained in tension. This procedure produces crazed specimens in which the craze fibrils can be characterized by a well-defined draw ratio, ₀. The films are then exposed to electron irradiation. This produces chemical crosslinks between the molecules, thus forming a crosslinked network. Subsequent heating of the film aboveT _g results in the entanglement network trying to retract to=1. The crosslink network, however, tries to maintain the. of the craze fibrils at ₀. The craze fibrils thus retract to Ferry's state of ease, _S, where the tension of the entanglement network is balanced by the compression of the crosslink network. Measurements of _s in crazes crosslinked and then healed confirm that a 25 to 50% loss of entanglement density in craze fibrils occurs, in agreement with theoretical predictions.     

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.     

Role of carbon dissolution during manganese oxidation in CO-CO2 Atmospheres

The role of carbon produced from the decomposition of CO-CO₂ gas mixtures traditionally is ignored during oxidation experiments, and only the oxygen potential established by the gas mixture is generally considered. Accordingly, to overcome this problem, a model which takes into account both carbon and oxygen dissolution into a p-type scale has been developed and tested. Two distinct cases of the model were tested at 1002°C. In the first case, variable CO/CO₂ ratios were used to determine the parabolic kinetics. Thus, both variable oxygen and carbon potentials were studied. In the p-type region of MnO, the developed model appeared to hold quite well. In the second case, constant CO-CO₂ ratios diluted with helium were utilized to maintain a constant oxygen potential with a variable carbon activity. The developed model appeared to hold again for the p-type region of MnO. Metallographically it was noted that the MnO scale developed exhibited an outer compact region and an inner layer that was somewhat porous.