One of the major sulphated proteins secreted by rat hepatocytes contains low-sulphated chondroitin sulphate.

When isolated hepatocytes are incubated with 35SO4(2-), a specific set of secretory proteins is labelled. One of these proteins is electrophoretically heterogeneous, with an apparent molecular mass of 35-45 kDa [Marcks von Würtemberg & Fries (1989) Biochemistry 28, 4088-4093]. Here we report that treatment with chondroitinase ABC converted the broad electrophoretic band of this protein, with a 50-60% loss of radioactivity, into a relatively homogeneous band with a molecular mass of 28 kDa. Size determination by gel chromatography of the protein's oligosaccharide chain (released by alkali treatment) indicated that it contained about 40 hexose units. Similar analysis of the enzyme-resistant oligosaccharide chain remaining linked to the protein after chondroitinase ABC treatment indicated a size of between six and eight hexose units. These observations suggest that the protein's oligosaccharide chain carries only three or four sulphate groups, of which one or two are located close to the polypeptide chain. Consistent with this hypothesis, the free oligosaccharide behaved like a low-sulphated glycosaminoglycan upon ion-exchange chromatography.     

An in vitro bioassay for quantitation of human interferons by measurements of antiproliferative activity on a continuous human lymphoma cell line

Interferons have, in addition to their antiviral effects, been shown to possess several non-antiviral activities. In this study, an in vitro bioassay for interferon alpha (IFN-alpha) preparations based on their antiproliferative effect in cultured Daudi cells has been developed. Briefly, about 10(5) cells per ml treated with different concentrations of IFN were incubated under standard culture conditions for 3 days. Two different end points, i.e. incorporation of [3H]thymidine and final cell density, were used and responses were evaluated according to established pharmacopoeial principles for quantification of biomolecules. Both methods gave similar results. However, measurement of final cell density yielded the most precise results. The proposed assay, with an effective assay range of 1-10 IU/ml (approximately 0.2-2 x 10(-12)M, had a high sensitivity and precision as well as a good reproducibility. Compared with antiviral assays, it is less resource demanding. In conclusion, the in vitro bioassay described is well suited for potency determinations of IFN-alpha and probably also IFN-beta preparations.     

Species composition of dabbling duck assemblages: ecomorphological patterns compared with null models

Ecomorphological patterns of breeding dabbling duck (Anas spp.) assemblages were studied in six regions in northern Europe. Observed spacings among species in terms of bill lamellar density and body length were compared with expected spacings based on null models incorporating different levels of constraints (regional species pools, species relative abundances, lake size and habitat requirements of species). Deviations of observed spacings from expected ones were compared with prey abundance and prey size diversity in the lakes. Observed spacings in terms of body length, but not in terms of bill lamellar density, were greater than expected on the basis of null models. The most abundant species were generally relatively more different than less abundant species in terms of body length but not in terms of bill lamellar density. Deviations between observed and expected spacings in terms of body length were more like those predicted by the competition hypothesis in lakes with low food abundance than in lakes with high food abundance. Patterns in bill lamellar spacings were not related to food abundance nor to food size diversity. In general, patterns in body length spacings were consistent with the competition hypothesis whether the null model used in comparisons was constrained or not.     

Psychological complications after stillbirth--influence of memories and immediate management: population based study.

OBJECTIVE--To identify factors that may predict long term psychological complications among women who have had a stillborn child. DESIGN--Nationwide population based study using epidemiological methods. SUBJECTS--380 subjects and 379 controls who had had a stillborn or non-deformed live child in Sweden in 1991. RESULTS--Information was provided by 636 (84%) women. The ratio (95% confidence interval) of proportions of women with symptoms related to anxiety above the 90th centile for women who had had a stillborn child compared with those who had not was 2.1 (1.2 to 3.9). An interval of 25 hours or more from the diagnosis of death in utero to the start of delivery gave a ratio of 4.8 (1.5 to 15.9). The ratio was 2.3 (1.1 to 5.3) for not seeing the child as long as the mother had wished and 3.1 (1.6 to 6.0) for no possession of a token of remembrance. CONCLUSION--It is advisable to induce the delivery as soon as feasible after the diagnosis of death in utero. A calm environment for the woman to spend as much time as she wants with her stillborn child is beneficial, and tokens of remembrance should be collected.     

Enhanced production of glycerol in an alcohol dehydrogenase (ADH I) deficient mutant of Saccharomyces cerevisiae

Summary A partial alcohol dehydrogenase, ADH I, deficient mutant, GRF 18-2 of S. cerevisiae has been isolated. The mutant is resistant to allyl alcohol and the spec. activity of ADH I is 15-fold reduced in the mutant. In a batch fermentation the mutant overproduces glycerol. The production is enhanced 6–7 fold compared with the wildtype strain and it amounts to about 40 per cent of the ethanol produced. The yield of ethanol and glycerol is 56 and 24 per cent respectively. Another mutant possibly defect in the gene for ADH II has a reduced capacity to oxidize ethanol.     

Complete remission in a patient with acute myelogenous leukemia treated with leukocyte α-interferon and cimetidine

Summary A 76-year-old woman with acute myelogenous leukemia with approximately 65% myeloblasts on bone marrow examination was treated daily with a combination of 4 megaU of leukocyte interferon IM and 1,000 mg cimetidine PO. During therapy there was a gradual decrease of bone marrow myeloblasts down to 9% and a normalization of peripheral white blood cells. The treatment was discontinued after 6 weeks because of increasing fatigue and anorexia. The general condition improved greatly during the following weeks and the patient entered complete remission, which has continued for 6 months so far. In the course of therapy there was a gradual appearance of antibodies showing a selective binding capacity to autochthonous leukemic cells with no tendency to increased binding to remission cells. The aim of this report is to stimulate a further evaluation of this form of therapy in additional AML patients whenever this might be justified as an alternative to conventional chemotherapy.     

Size and shape of two intestinal dipeptidases

Physicochemical parameters were determined on glycyl-L-leucine hydrolase (glycy-leucine dipeptidase, EC 3.4.13.2) and aminoacyl-L-proline hydrolase (proline dipeptidase, EC 3.4.13.9), purified from pig small intestine. The native molecular weights were found to be 115,000 and 113,000, respectively, as determined by a sedimentation equilibrium technique. Under denaturing conditions the molecular weights were found to be 51,000 and 63,200, respectively, using the same technique. It is concluded that each dipeptidase is composed of two subunits of equal molecular weight. The two dipeptidases have the same Stokes radius, 4.2 nm, analysed by gel chromatography. The sedimentation coefficients were found to be 5.8. S and 6.5 S and the intrinsic viscosities 5.4 ml/g and 5.8 ml/g, respectively. For both dipeptidases the measured physicochemical parameters are in accordance with the model of a prolate ellipsoid of revolution, having an axial ratio of about 5.     

Periodate oxidation and alkaline degradation of heparin-related glycans

Heparin, heparan sulphate, and various derivatives thereof have been oxidised with periodate at pH 3.0 and 4° and at pH 7.0 and 37°. Whereas oxidation under the latter conditions destroys all of the nonsulphated uronic acids, treatment with periodate at low pH and temperature causes selective oxidation of uronic acid residues. The reactivity of uronic acid residues depends on the nature of neighbouring 2-amino-2-deoxyglucose residues. d-Glucuronic acid residues are susceptible to oxidation when flanked by N-acetylated amino sugars, but resistant when adjacent residues are either unsubstituted or N-sulphated. L-Iduronic acid residues in their natural environment (2-deoxy-2-sulphoamino-d-glucose) are resistant to oxidation, whereas removal of N-sulphate groups renders a portion of these residues periodate-sensitive. Oxidised uronic acid residues in heparin-related glycans may be cleaved by alkali, producing a series of oligosaccharide fragments. Thus, periodate oxidation-alkaline elimination provides an additional method for the controlled degradation of heparin.     

Epidemiological investigation of an animal house based upon phage-typing and biotyping of Staphylococcus epidermidis.

The value of biotyping and phage-typing coagulase-negative staphylococci in the epidemiological investigation of a laboratory animal house was clearly demonstrated. In the animal rooms in which conventional bacteriological methods revealed equal bacterial contamination between a conventional unit and one housing specified-pathogen-free rodents, biotyping identified Staphylococcus cohnii as the only species in the latter, compared to S. warneri, S. hominis, S. saprophyticus. S. xylosus abd S. epidermidis as well as S. cohnii in the conventional unit. Similarly, phagetyping revealed 2 phage types in the specified-pathogen-free compared to 7 in the conventional unit. Thus biotyping and phage-typing provided evidence for the existence of a barrier between these units that had presented similar gross bacteriological findings.