N-glycosylation of recombinant human interferon-gamma produced in different animal expression systems

Recombinant human interferon-gamma (IFN-gamma) was expressed in Chinese hamster ovary cells, baculovirus-infected Sf9 insect cells and the mammary gland of transgenic mice. The N-linked carbohydrate populations associated with both Asn25 and Asn97 glycosylation sites were characterized by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) in combination with exoglycosidase array sequencing. A site-specific analysis of dual (2N) and single (1N) site-occupancy variants of IFN-gamma derived from Chinese hamster ovary cells showed that N-glycans were predominantly of the complex bi- and triantennary type. Although Asn25-linked glycans were substituted with a core fucose residue, Asn97 N-glycans were predominantly non-fucosylated, and truncated complex and high-mannose oligosaccharide chains were also evident. Transgenic mouse derived IFN-gamma exhibited considerable site-specific variation in N-glycan structures. Asn25-linked carbohydrates were of the complex, core fucosylated type, Asn97-linked carbohydrates were mainly of the oligomannose type, with smaller proportions of hybrid and complex N-glycans. Carbohydrates associated with both glycosylation sites of IFN-gamma from Sf9 insect cells were mainly tri-mannosyl core structures, with fucosylation confined to the Asn25 site. These data demonstrate the profound influence of host cell type and protein structure on the N-glycosylation of recombinant proteins.     

Treatment of wastewater containing Pb and Fe using ion‐exchange techniques

Treatment of wastewater containing lead and iron was examined using two different ion‐exchange resins namely Duolite ES 467 (containing amino‐phosphonic functional groups) and a chelating ion‐exchange resin (containing hydroxamic acid functional groups). Initially different sorption parameters such as contact time, pH, concentrations of sorbent, sorbate and chloride ion were studied. The sorption kinetics was observed to be fast and equilibrium could be reached within 30 min. Lead sorption efficiency increased with increase in pH whereas the opposite trend was observed with iron. The presence of chloride ions greatly reduced the Pb sorption efficiency in the case of Duolite ES 467. Column studies were carried out to recover Pb and Fe individually using Duolite ES 467 resin. The maximum uptake of Pb at pH 2 and 3 was observed to be 11.63 and 33.96 g dm^?3 of resin respectively. Similarly, for Fe at pH 2 and 3 the uptake was observed to be 10.07 and 6.96 g dm^?3 of resin respectively. In the presence of chloride ions, column studies were carried out using Duolite ES 467 for iron and chelating ion‐exchange resin containing hydroxamic acid functional groups for lead sorption. Hydroxamic acid resin's loading capacity remains constant for at least up to 20 cycles. Copyright © 2005 Society of Chemical Industry     

The use of trypsin and lysozyme immobilized on a cellulose carrier in treating otitis externa

In December 1991, an outbreak of Streptococcus bovis septicaemia occurred in a Belgian pigeon loft where 25 male and 25 female racing pigeons were housed. The main clinical signs included inability to fly and poor breeding results. None of the female pigeons and only one male pigeon was able to fly. Nine affected pigeons were necropsied. Histologically a tenosynovitis of the tendon of the deep pectoral muscle was observed in most of them and S bovis was isolated from the canalis triosseus or the shoulder joints of five of the nine pigeons. The pigeons were successfully treated with ampicillin administered in the drinking water for seven days.     

Further evidence for elevated human minisatellite mutation rate in Belarus eight years after the Chernobyl accident

Analysis of germline mutation rate at human minisatellites among children born in areas of the Mogilev district of Belarus heavily polluted after the Chernobyl accident has been extended, both by recruiting more families from the affected region and by using five additional minisatellite probes, including multi-locus probe 33.6 and four hypervariable single-locus probes. These additional data confirmed a twofold higher mutation rate in exposed families compared with non-irradiated families from the United Kingdom. An elevated rate was seen at all three independent sets of minisatellites (detected separately by multi-locus probes 33.15, 33.6 and six single-locus probes), indicating a generalised increase in minisatellite germline mutation rate in the Belarus families. Within the Belarus cohort, mutation rate was significantly greater in families with higher parental radiation dose estimated for chronic external and internal exposure to caesium-137, consistent with radiation induction of germline mutation. The spectra of mutation seen in the unexposed and exposed families were indistinguishable, suggesting that increased mutation observed over multiple loci arises indirectly by some mechanism that enhances spontaneous minisatellite mutation.     

Frequency of chromosome disorders occurring in human lymphocytes in the in vitro exposure to different doses of deuterons as protected by adeturon

Peripheral blood of 4 healthy donors was exposed to deutrons with the energy 4,2 GeV/nuclone and the intensivity of 4 to 6.10(10) particles per impulse, the capacity of the dose being 0,008 Gr/sec. The LPE of deutrons was 2,14 MeV sm2/g. Irradiation doses were 0,26; 0,49; 0,97; 2,02; 3,80 Gr. 15-20 min prior to irradiation, the protector adeturone was added at a concentration of 550 micrograms/ml of blood. Chromosome aberrations were analyzed in protected and unprotected blood samples. The results were treated using the method of variance and regression analysis. The correlative correspondence between the number of chromosome aberrations and an approximate dose of actual irradiation was established as Y = 0,13 d + 6,55.10(-4) D2 for dicentrics and as 0,26 D + 12,6.10(-4) D2 + 4,5 for the total percentage of aberrations. The pattern of the dose - effect dependence was retained in protected samples. The calculated values of the factor of dose decrease confirmed antiirradiation properties of adeturone.