Observations on the influence of glutamine,asparagine and peptone on growth and t-PA production of Chinese hamster ovary (CHO) cells

When a transfected CHO cell, that produces tissue-type Plasminogen Activator, t-PA, was transferred from a medium based on 5% Fetal Calf Serum, FCS, to a medium based on 0.8% casein peptone with variable glutamine and asparagine content, it was observed, that the growth of the cells changed from anchorage dependant to suspension culture giving more reproducible cultivations. In the FCS culture t-PA was unstable, observed as a decline in t-PA concentration after 250 h. This decline in t-PA concentration was not observed in the serum free culture, although there was a decline in productivity after 200 h. This change in production profile may be attributed to either no proteolytic attack from serum or by scavenging of proteolytic activities produced by the cells from the peptone peptides. Increasing amounts of glutamine/asparagine gave higher production of t-PA in synchrony with an increasing production of ammonia/ammonium ions. Ammonia inhibition does not seem to be a key factor for this cell line as seen with many others.     

The genus Ptyssiglottis (Acanthaceae). A taxonomic monograph

A taxonomic treatment of the genus Ptyssiglottis including Ancylacanthus, Hallieracantha, Oreothyrsus , and Polytrema is given. The phylogeny is briefly dealt with. Leaf morphology, inflorescence morphology and pollen morphology yield important characters. The distribution of selected characters is mapped. The distribution of all species is mapped. Eight new species are published viz. P. campanulata, P. decurrens, P. fusca, P. glandulifera, P. longisepala, P. mucronata, P. pubescens , and P. stamino-difera . Fifteen new combinations are made viz. P. caudata, P. creaghii, P. cuprea, P. cyrtandroides, P. dulcamarioides, P. fastidiosa, P. glabrisepala, P. granulata, P. nigrescens, P. peranthera, P. psychotriifolia, P. pubisepala, P. salicifolia, P. sanguinolenta , and P. undulata .     

Mallomonas palaestrica sp. nov. (Synurophyceae), a new member of sect. Torquatae from Denmark

Mallomonas paluestrica sp. nov., a new member of sect. Torquatae ser. Pumilae , was found in a small Danish pond and is described and illustrated with TEM and SEM micrographs.     

Migratory behaviour and growth of hatchery-reared post-smolt Atlantic salmon Salmo salar

Individually lagged, 1+ and 2+ hatchery-reared smolts of Atlantic salmon were released in spring and early summer at the mouth of the R. Imsa, south-western Norway. The post-smolts moved mainly northwards in the sea with the coastal current. The estimated mean migratory speed (± s.d. ) of those captured in the sea along the Norwegian coast was 7.45 (± 6.26) km day ⁻¹; in the fjords it was 1.63 (± 2.33) km day⁻². Many of the post-smohs ascended rivers the same year as released; 37.3% of the total number recaptured were caught in R. Imsa, upstream from the site of release, and 5.8% were caught in other rivers throughout middle and southern parts of Norway. The fish recaptured in rivers was probably sexually mature and entered rivers to spawn. Mean specific growth rate for post-smolts caught in the sea was higher than for those caught in R. Imsa (P <0.001) but not for those caught in other rivers (P> 0.05). Post-smolts ascending R. Imsa were smaller at release than those ascending other rivers. However, there was no size difference at release between post-smolts captured in the sea and those recaptured in rivers other than the R. Imsa.     

Insertion of Transposon Tn917 Derivatives into the Lactococcus lactis subsp. lactis Chromosome

Two transposition vectors, pTV32 and pLTV1, containing transposon Tn917 derivatives TV32 and LTV1, respectively, were introduced into Lactococcus lactis subsp. lactis MG1614. It was found that pTV32 and pLTV1 replicate and that TV32 and LTV1 transpose in this strain. A protocol for production of a collection of Tn917 insertions in L. lactis subsp. lactis was developed. The physical locations of TV32 on the chromosomal SmaI fragments of 62 independent transpositions were established by pulsed-field gel electrophoresis. These transpositions could be divided into at least 38 different groups that exhibited no Tn917-dominating hot spots on the L. lactis subsp. lactis chromosome. A total of 10 of the 62 transpositions resulted in strains that express β-galactosidase. This indicates that there was fusion of the promoterless lacZ of the Tn917 derivatives to a chromosomal promoter. Thus, the Tn917-derived transposons should be powerful genetic tools for studying L. lactis subsp. lactis.     

Isolation and characterization of an extracellular glycosylated protein complex from Clostridium thermosaccharolyticum with pectin methylesterase and polygalacturonate hydrolase activity.

An extracellular protein complex was isolated from the supernatant of a pectin-limited continuous culture of Clostridium thermosaccharolyticum Haren. The complex possessed both pectin methylesterase (EC 3.1.1.11) and exo-poly-alpha-galacturonate hydrolase (EC 3.2.1.82) activity and produced digalacturonate from the nonreducing end of the pectin chain. The protein consisted of 230- and 25-kDa subunits. The large subunit contained 10% (wt/wt) sugars (N-acetylgalactosamine and galactose). Under physiological conditions both activities acted in a coordinated manner: the ratio between methanol and digalacturonate released during degradation was constant and equal to the degree of esterification of the pectin used. Prolonged incubation of the enzyme with pectin led to a nondialyzable fraction that was enriched in neutral sugars, such as arabinose, rhamnose, and galactose; the high rhamnose/galacturonic acid ratio was indicative of hairy region-like structures. The smallest substrate utilized by the hydrolase was a tetragalacturonate. Vmax with oligogalacturonates increased with increasing chain length. The Km and Vmax for the polygalacturonate hydrolase with citrus pectate as a substrate were 0.8 g liter-1 and 180 mumol min-1 mg of protein-1, respectively. The Km and Vmax for the esterase with citrus pectin as a substrate were 1.2 g liter-1 and 440 mumol min-1 mg of protein-1, respectively. The temperature optima for the hydrolase and esterase were 70 and 60 degrees C, respectively. Both enzyme activities were stable for more than 1 h at 70 degrees C. The exo-polygalacturonate hydrolase of Clostridium thermosulfurogenes was partially purified while the methylesterase was also copurified.     

Anaerobic degradation of dimethylsulfoniopropionate to 3-S-methylmercaptopropionate by a marine Desulfobacterium strain

Dimethylsulfoniopropionate, an osmolyte of marine algae, is thought to be the major precursor of dimethyl sulfide, which plays a dominant role in biogenic sulfur emission. The marine sulfate-reducing bacterium Desulfobacterium strain PM4 was found to degrade dimethylsulfoniopropionate to 3-S-methylmercaptopropionate. The oxidation of one of the methyl groups of dimethylsulfoniopropionate was coupled to the reduction of sulfate; this process is similar to the degradation betaine to dimethylglycine which was described earlier for the same strain. Desulfobacterium PM4 is the first example of an anaerobic marine bacterium that is able to demethylate dimethylsulfoniopropionate.Abbreviations DMSP dimethylsulfoniopropionate - DMS dimethyl sulfide - MMPA 3-S-methylmercaptopropionate     

Phenotypic effects of overexpression of Agrobacterium rhizogenes T-DNA ORF13 in transgenic tobacco plants are mediated by diffusible factor(s)

Nicotiana tabacum cv. Xanthi transgenic plants expressing ORF13 of Agrobacterium rhizogenes 8196 T-DNA under the 35S RNA promoter from the cauliflower mosaic virus displayed developmental abnormalities. They were small, with short and variable internodal lengths, their root systems were poorly developed; leaves were small, asymmetric, rounded, wrinkled and dark green; flowers were short, and irregularly shaped. They exhibited reduced apical dominance and regularly produced offshoots at the base of the plant. This phenotype was also exhibited by offshoots of normal N. tabacum cv. Xanthi stock grafted with a transgenic scion indicating that expression of ORF13 influences plant development via diffusible factor(s).     

Heterostyly in the Canarian endemic Jasminum odoratissimum (Oleaceae)

Jasminum odoratissimum is a Madeira and Canary Islands endemic showing classic heterostyly, i.e. with long-styled flowers with anthers at a low level in the corolla tube and short-styled flowers with anthers at a high level in the corolla tube. Short-styled flowers have large pollen, whereas long-styled flowers have small pollen. The two types are present in equal frequencies in the population.     

On-line membrane inlet mass spectrometry for feed-back control of precursor concentration in penicillin fermentation

Summary A sampling system which enables on-line measurements of the precursor phenoxyacetic acid (POAA) in penicillin fermentation by membrane inlet mass spectrometry is presented and its capacity for feed-back regulation of POAA to a low predefined concentration in a penicillin-V fermentation over 150 hours is demonstrated. The system measures alternately filtered sample and standard solution in a measuring cycle which is shorter than the response time of membrane inlet mass spectrometry (MIMS) but sufficiently long to decide whether the concentration of POAA in the sample is higher or lower than in the standard solution at set point concentration. The decision is used for on-off regulation of the addition of POAA.