The membrane dialysis bioreactor with integrated radial-flow fixed bed —a new approach for continuous cultivation of animal cells

A hybridoma cell was cultivated continuously in a membrane dialysis bioreactor with an integrated radial-flow fixed bed consisting of porous Siran^® carriers over a period of 6 weeks. Antibodies accumulated to an average of 100 mg l^?1, approx. 10 times more than in fixed bed cultures without dialysis membrane. Serum costs could be reduced about 85% due to an appropriate feeding strategy. Siran^® carriers with 3–5 mm diameter showed an advantage compared to those with 1–2 mm diameter. For the 3–5 mm carrier the specific glucose uptake rate and the MAb production rate were constant, if the velocity was between 0.09 mm s^?1 and 0.75 mm s^?1. At higher velocities cells are washed out of the bed. Furthermore antibody consistency and cell stability were verified in long-term cultivations over a period of 96 days. From an estimation of the antibody concentration reachable with the reactor concept under optimal conditions a concentration 45 times higher compared to axial-flow fixed bed reactors and 11 times higher compared to stirred tank reactors can be expected.     

Induction of transverse polarity by blue light: An all-or-none response

Phototropic stimulation induces a spatial memory. This was inferred from experiments with maize (Zea mays L.) coleoptiles involving opposing blue-light pulses, separated by variable time intervals, and rotation on a horizontal clinostat (Nick and Schafer, 1988b, Planta 175, 380-388). In those experiments, individual seedlings either curved towards the first or towards the second pulse, or they remained straight. Bending, if it occurred, seemed to be an all-or-none response. Intermediates, i.e. plants, bending only weakly, were not observed. In the first part of the present study it was attempted to create such intermediates. For this purpose the strength of the first, inducing, and the second, opposing, pulse was varied. The result was complex: (i) Individual seedlings maintained the all-or-none expression of spatial memory. (ii) However, on the level of the whole population, the time intervals at which a given response type dominated depended on the fluence ratio. (iii) Furthermore, the final curvature was determined by the fluence ratio. These results are discussed in terms of a blue-light-induced transverse polarity. This polarity initiates from a labile precursor, which can be reoriented by an opposing stimulation (indicated by the strong bending towards the second pulse). The strong curvatures towards the first pulse over long time intervals reveal that, eventually, the blue-light-induced transverse polarity becomes stabilised and thus immune to the counterpulse. In the second part of the study, the relation between phototropic transduction and transverse polarity was characterised by a phenomenological approach involving the following points: (i) Sensory adaptation for induction of transverse polarity disappears with a time course similar to that for phototropic sensory adaptatation. (ii) The fluence response for induction of transverse polarity is a saturation curve and not bell-shaped like the curve for phototropism (iii) For strong counterpulses and long time intervals the clinostat-elicited nastic response (Nick and Schafer 1989, Planta 179, 123-131) becomes manifest and causes an "aiming error" towards the caryopsis. (iv) Temperature-sensitivity of polarity induction was high in the first 20 min after induction, then dropped sharply and rose again with the approach of polarity fixation. (v) Stimulus-summation experiments indicated that, for different inducing fluences, the actual fixation of polarity happened at about 2 h after induction. These experiments point towards an early separation of the transduction chains mediating phototropism and transverse polarity, possibly before phototrophic asymmetry is formed.     

Chromosomal Integration and Expression of Two Bacterial alpha-Acetolactate Decarboxylase Genes in Brewer's Yeast

A bacterial gene encoding alpha-acetolactate decarboxylase, isolated from Klebsiella terrigena or Enterobacter aerogenes, was expressed in brewer's yeast. The genes were expressed under either the yeast phosphoglycerokinase (PGK1) or the alcohol dehydrogenase (ADH1) promoter and were integrated by gene replacement by using cotransformation into the PGK1 or ADH1 locus, respectively, of a brewer's yeast. The expression level of the alpha-acetolactate decarboxylase gene of the PGK1 integrant strains was higher than that of the ADH1 integrants. Under pilot-scale brewing conditions, the alpha-acetolactate decarboxylase activity of the PGK1 integrant strains was sufficient to reduce the formation of diacetyl below the taste threshold value, and no lagering was needed. The brewing properties of the recombinant yeast strains were otherwise unaltered, and the quality (most importantly, the flavor) of the trial beers produced was as good as that of the control beer.     

Neutralizing effect of zinc oxide on dehydroabietic acid-induced toxicity on human polymorphonuclear leukocytes

The cytotoxic effect of dehydroabietic acid (DHAA), a resin acid found in rosin, was studied on human polymorphonuclear leukocytes using leakage of 51Cr from prelabeled cells, supravital staining, and transmission electron microscopy. DHAA caused a strong dose-related release of 51Cr, a high uptake of trypan blue, and total cell necrosis, as seen in transmission electron microscopy. Albumin slightly reduced the toxic effects, whereas the addition of zinc in various forms strongly inhibited these toxic effects of DHAA in the concentration range of 10-500 micrograms/mL. In the presence of albumin, zinc oxide as a suspension inhibited the damage of the cell membranes more than a filtrate of zinc oxide, indicating a subsequent slow release of zinc from the zinc oxide.     

Environmental factors affecting the life-tables ofTetranychus urticae (Acari: Tetranychidae) III. Host-plant nutrition

First, the literature of the last two decades on nutritional effects of host plants on spider mites is briefly reviewed. Second, experiments are described that subjected micro-propagated apple trees to four different levels of each macronutrient N, P and K. Spider mites (Tetranychus urticae Koch) feeding on leaf disks of these plants were checked for their developmental time, egg production and longevity. Plant analysis revealed that the concentration of N, P and K corresponded to the respective treatments. The content of phenolic compounds in the leaves increased with N and P deficiency. In the N experiment, spider-mite preimaginal developmental rate and oviposition rate were both positively correlated with leaf N. Often, fecundity was positively correlated with N and carbohydrate content of the leaves, and negatively with the phenolic content. Longevity of the two-spotted spider mite was not significantly affected by any treatment. The K experiments yielded only minor differences in plant contents as well as in spider-mite biology.From these mite data, file-tables were constructed and statistically analyzed by the Jackknife technique. The life-table analysis showed a gradual decline in the intrinsic rate of natural increase (r _m) with N and P deficiency. With all experiments pooled,r _m was clearly correlated to leaf N and particularly to the content of phenolic compounds in the leaves. Nitrogen shortage had the most distinct influence on mite population growth: in a range of 1.5–3.0% leaf N,r _m increased by a factor of 4, the number of multiplications per generation (R ₀) by 11, and the doubling time of the population was prolonged 4-fold on severely N deficient leaves.     

Investigations on possible genotoxic effects ofFusarium toxins in boars

For several weeks, boars were fed feedstuff containing mycotoxins (zearalenone, nivalenol, and deoxynivalenol). To determine a possible mutagenic effect of this feedstuff, the boars were examinated for structural chromosome aberrations in the lymphocytes. The investigations indicate a genotoxic effect on the boars’ lymphocytes.     

Responses of the photosynthetic flagellate,Euglena gracilis,to hypergravity

Motility and orientation has been studied in the unicellular photosynthetic flagellate, Euglena gracilis, using real time image analysis capable of tracking up to 200 cells simultaneously in the slow rotating centrifuge microscope (NIZEMI) which allows one to observe the cells' swimming behavior during centrifugation accelerations between 1 g and 5 g. At 1 g the cells show a weak negative gravitaxis, which increases significantly at higher accelerations up to about 3 g. Though most cells were capable of swimming even against an acceleration of 4.5 g, the degree of gravitaxis decreased and some of the cells were passively moved downward by the acceleration force; this is true for most cells at 5 g. The velocity of cells swimming against 1 g is about 10% lower than that of cells swimming in other directions. The velocity decreases even more drastically in cells swimming against higher acceleration forces than those at 1 g. The degree of gravitactic orientation drastically decreases after short exposure to artificial UV radiation which indicates that gravitaxis may be due to an active physiological perception rather than a physical effect such as an asymmetry of the center of gravity within the cell. Offprint requests to: D.-P. Häder