Effects of bubble–liquid two‐phase turbulent hydrodynamics on cell damage in sparged bioreactor

According to recent experimental studies on sparged bioreactors, significant cell damage may occur at the gas inlet region near the sparger. Although shear stress was proposed to be one of the potential causes for cell damage, detailed hydrodynamic studies at the gas inlet region of gas–liquid bioreactors have not been performed to date. In this work, a second‐order moment (SOM) bubble–liquid two‐phase turbulent model based on the two‐fluid continuum approach is used to investigate the gas–liquid hydrodynamics in the bubble column reactor and their potential impacts on cell viability, especially at the gas inlet region. By establishing fluctuation velocity and bubble–liquid two‐phase fluctuation velocities correlation transport equations, the anisotropy of two‐phase stresses and the bubble–liquid interactions are fully considered. Simulation results from the SOM model indicate that shear and normal stresses, turbulent energy dissipation rate, and the turbulent kinetic energy are generally smaller at the gas inlet region when compared with those in the fully developed region. In comparison, a newly proposed correlation expression, stress‐induced turbulent energy production (STEP), is found to correlate well with the unusually high cell death rate at the gas inlet region. Therefore, STEP, which represents turbulent energy transfer to a controlled volume induced by a combination of shear and normal stresses, has the potential to provide better explanation for increased cell death at the sparger region. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:48–58, 2014     

Dual aerobic hollow-fiber bioreactor for cultivation of Streptomyces aureofaciens

Streptomyces aureofaciens (ATCC 12416c) was grown in the interstitial region formed by a parallel arrangement of three hollow silicone tubules contained within a microporous polypropylene hollow fiber. Liquid-soluble nutrients were supplied by diffusion across the polypropylene fiber to the interstitial cell-containing region whereas air or oxygen was provided by diffusion from the silicone tubule lumina to the cell mass. In this bioreactor, S. aureofaciens grew to high cell densities (greater than 10(11) cells/cm(3)) and the culture so-obtained continously synthesized the secondary metabolite tetracycline. The volumetric productivity of tetracycline based on the interstitial volume was 90 mug/ml/h and based on the total reactor volume was 5.5 mug/mL/h. The high surface area-to-volume ratio afforded by the cylindrical configuration together with spatially distinct conduits to continuously transport liquids and gases, each of which may be nutrients or products of biosynthesis, to or from a tissuelike cell mass provides an alternative to the conventional air- or oxygen-sparged fermentation vessel. High volumetric reactor productivities may be achieved by virute of the concentrated stationary cell mass and by the appropriate selection of fiber sizes and materials so as to ensure adequate supplies of liquid and gaseous substrates to, as well as removal of metabolites from, most cells in the culture. This reactor topology is quite general and may be adapted to most microbial as well as mammalian and plant cell systems.     

Cell physiology and antibiotic production of Streptomyces coelicolor grown on solid medium

Summary In order to examine the physiology ofStreptomyces coelicolor when growing on solid media, we have employed a membrane overlay technique and used a new approach to extract substrate and product compounds from the agar. Comparisons made with liquid grown cultures indicate a change from non-growth associated productivity of actinorhodin in liquid culture, to growth associated production on agar plates. In contrast, the temporal control of methylenomycin production was virtually identical under both culture conditions. Considerable extracellular protein production was observed during growth on agar.     

Tylosin production by Streptomyces fradiae using raw cornmeal in airlift bioreactor

Using a 50-l airlift bioreactor, for the effective production of tylosin from Streptomyces fradiae TM-224 using raw cornmeal as the energy source, various environmental factors were studied in flask cultures. The maximum tylosin concentration was obtained at 32 degrees C and pH between 7.0 and 7.5. When seed was inoculated after 24 h of culture, the maximum tylosin concentration, 5.7 g/l, was obtained after 4 days of culture. Various concentrations of raw cornmeal were tested to investigate the optimum initial concentration for the tylosin production. An initial raw cornmeal concentration of 80 g/l gave the highest tylosin concentration, 5.8 g/l, after 5 days of culture. Of the various nitrogen sources, soybean meal and fish meal were found to be the most effective for the production of tylosin. In particular, with the optimal mixing ratio, 12 g/l of soybean meal to 14 g/l of fish meal, 7.2 g/l of tylosin was obtained after 5 days of culture. To compare raw cornmeal and glucose for the production oftylosin in the 50-1 airlift bioreactor for 10 days, fed-batch cultures were carried out under the optimum culture conditions. When raw corn meal was used as the energy source, the tylosin production increased with increasing culture time. The maximum tylosin concentration after 10 days of culture was 13.5 g/l, with a product yield from raw cornmeal of 0.123 g/g of consumed carbon source, which was about 7.2 times higher than that obtained when glucose was used as the carbon source.     

Effects of sugars on the physiology of cultured fibroblasts

Pronounced, rapid, and reversible changes in cultured normal and transformed fibroblasts are observed following removal of sugars from the culture medium. Growth immediately ceases or greatly diminishes; it resumes at a normal rate without any appreciable delay following re-addition of glucose. The cell shape changes within 6 h to a more flattened and elongated form similar to that of cells treated with db-cAMP. The NAD⁺/NADH ratio rises 4-fold and the ATP level falls about 25% in 2 h; however, the cAMP level remains constant. In transformed mouse SVT2 cells, other sugars will not replace glucose. A variant line of SVT2 (SVT2 SUG⁺) which will grow on numerous sugars has been isolated. Its growth rate, morphology and adhesion to the substratum, but not cAMP content, are dependent upon the sugar composition. These results suggest that the sugar composition of the growth medium has pronounced effects on cell physiology and that the effects are not mediated by cAMP.     

Effects of substratum morphology on cell physiology

Among the host of substratum properties that affect animal cell behavior, surface morphology has received relatively little attention. The earliest effect of surface morphology on animal cells was discovered almost a century ago when it was found that cells became oriented in response to the underlying topography. This phenomenon is now commonly known as contact guidance. From then until very recentrly, little progress has been made in understanding the role of surface morphology on cell behavior, primarily due to a lack of defined surfaces with uniform morphologies. This problem has been solved recently with the development of photolithographic techniques to prepare substrata with well defined and uniform surface morphologies. Availability of such surfaces has facilitated systematic in vitro experiments to study influence of surface morphology on diverse cell physiological aspects such as adhesion, growth, and function. For example, these studies have shown that surfaces with uniform multipls parallel grooves can enhance cell adhesion by confining cells in grooves and by mechanically interlocking them. Several independent studies have demosterated that cell shape is a major determinant of cell growth and function. Because surface morphology has been shown to modulate the extent of cell spreading and cell shape, its effects on cell growth and function appear to be mediated via this biological coupling between cell shape and function. New evidence in the cell biology literature is emerging to suggest that surface morphology could affect other cell behavioral properties such as post-translational modifications. Further elucidation of such effects will enable better designs for implant and cell culture substrata.     

Growth of Streptomyces hygroscopicus in rotating-wall bioreactor under simulated microgravity inhibits rapamycin production

Growth of Streptomyces hygroscopicus under conditions of simulated microgravity in a rotating-wall bioreactor resulted in a pellet form of growth, lowered dry cell weight, and inhibition of rapamycin production. With the addition of Teflon beads to the bioreactor, growth became much less pelleted, dry cell weight increased but rapamycin production was still markedly inhibited. Growth under simulated microgravity favored extracellular production of rapamycin, in contrast to a greater percentage of cell-bound rapamycin observed under normal gravity conditions. Received: 20 September 1999 / Received revision: 18 November 1999 / Accepted: 19 November 1999     

Effects of mist irrigation on the physiology of sugar beet

Bacilliform particles characteristic of plant rhabdoviruses were found in negatively-stained leaf sap and in thin sections of Laburnum anagyroides in England showing vein yellowing. The particles were detected principally in the perinuclear space of parenchyma cells. They were not transmitted by sap inoculation to twelve herbaceous species. The affected trees also showed mosaic symptoms but there was no evidence of an association between these and the bacilliform particles.     

Effects of serotonin on the physiology of the rabbit small intestine

Serotonin has been shown to alter the intestinal transport of ions and intestinal motility. These effects may interfere with each other, modulating the whole physiology of the intestine. We have previously shown that serotonin also alters the transport of nutrients. Thus, the aims of the present work were to determine the possible interference between the secretagogue effect of serotonin and the mechanism by which serotonin inhibits the absorption of nutrients, and to study the effect of serotonin on the digestive activity of nutrients of the brush border membrane jejunum enterocyte in the rabbit. The results show that the secretagogue effect of serotonin neither affects the inhibitory effect of serotonin on the intestinal absorption of the nutrients, nor affects the activity of Na+/K+-ATPase. The activity of sucrase and aminopeptidase N was also not affected by serotonin in the rabbit jejunum. Finally, we also studied different parameters of the motility in the rabbit small intestine. Serotonin seemed to stimulate the motility of the rabbit small intestine by increasing integrated mechanical activity and tone of muscle fibers in duodenum, jejunum, and ileum. In conclusion, serotonin might alter or modulate the whole intestinal physiology.     

Pleiotropic role of the Sco1/SenC family copper chaperone in the physiology of Streptomyces

Antibiotic production and cell differentiation in Streptomyces is stimulated by micromolar levels of Cu²⁺. Here, we knocked out the Sco1/SenC family copper chaperone (ScoC) encoded in the conserved gene cluster ‘sco’ (the S treptomycesco pper utilization) in Streptomyces coelicolor A3(2) and S. griseus. It is known that the Sco1/SenC family incorporates Cu²⁺ into the active centre of cytochrome oxidase (cox). The knockout caused a marked delay in antibiotic production and aerial mycelium formation on solid medium, temporal pH decline in glucose‐containing liquid medium, and significant reduction of cox activity in S. coelicolor. The scoC mutant produced two‐ to threefold higher cellular mass of the wild type exhibiting a marked cox activity in liquid medium supplied with 10 µM CuSO₄, suggesting that ScoC is involved in not only the construction but also the deactivation of cox. The scoC mutant was defective in the monoamine oxidase activity responsible for cell aggregation and sedimentation. These features were similarly observed with regard to the scoC mutant of S. griseus. The scoC mutant of S. griseus was also defective in the extracellular activity oxidizing N,N′‐dimethyl‐p‐phenylenediamine sulfate. Addition of 10 µM CuSO₄ repressed the activity of the conserved promoter preceding scoA and caused phenylalanine auxotrophy in some Streptomyces spp. probably because of the repression of pheA; pheA encodes prephenate dehydratase, which is located at the 3′ terminus of the putative operon structure. Overall, the evidence indicates that Sco is crucial for the utilization of copper under a low‐copper condition and for the activation of the multiple Cu²⁺‐containing oxidases that play divergent roles in the complex physiology of Streptomyces.     

Effects of Metals on a Range of Streptomyces Species

A plate diffusion method was used to assess the tolerance of 28 mesophilic and 4 thermophilic species of streptomycetes to eight metals. This method allowed a qualitative screen of the effects of the metals on growth, on sporulation, and in some cases, on secondary metabolite production. Tolerant and sensitive species were identified, some of which exhibited the same response (i.e., tolerance or sensitivity) to a number of metals. Experiments in liquid culture were used to quantify the precise concentration ranges of the metals at which the most tolerant or sensitive species could grow. Distribution profiles of the sensitivities of all species examined toward different metals revealed that the order of toxicity was Hg > Cd > Co > Zn > Ni > Cu > Cr > Mn.     

Effects of 3,4-methylenedioxymethamphetamine administration on retinal physiology in the rat

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) is known to produce euphoric states, but may also cause adverse consequences in humans, such as hyperthermia and neurocognitive deficits. Although MDMA consumption has been associated with visual problems, the effects of this recreational drug in retinal physiology have not been addressed hitherto. In this work, we evaluated the effect of a single MDMA administration in the rat electroretinogram (ERG). Wistar rats were administered MDMA (15 mg/kg) or saline and ERGs were recorded before (Baseline ERG), and 3 h, 24 h, and 7 days after treatment. A high temperature (HT) saline-treated control group was also included. Overall, significantly augmented and shorter latency ERG responses were found in MDMA and HT groups 3 h after treatment when compared to Baseline. Twenty-four hours after treatment some of the alterations found at 3 h, mainly characterized by shorter latency, tended to return to Baseline values. However, MDMA-treated animals still presented increased scotopic a-wave and b-wave amplitudes compared to Baseline ERGs, which were independent of temperature elevation though the latter might underlie the acute ERG alterations observed 3 h after MDMA administration. Seven days after MDMA administration recovery from these effects had occurred. The effects seem to stem from specific changes observed at the a-wave level, which indicates that MDMA affects subacutely (at 24 h) retinal physiology at the outer retinal (photoreceptor/bipolar) layers. In conclusion, we have found direct evidence that MDMA causes subacute enhancement of the outer retinal responses (most prominent in the a-wave), though ERG alterations resume within one week. These changes in photoreceptor/bipolar cell physiology may have implications for the understanding of the subacute visual manifestations induced by MDMA in humans.     

Fine structure, physiology and biochemistry of arthrospore germination in Streptomyces antibioticus.

During germination, Streptomyces antibioticus arthrospores passed through stages: darkening, swelling and germ tube emergence. The first stage, darkening, whose main features were a decrease in absorbance and a loss of refractility, only required exogenous divalent cations (Ca2+, Mg2+ or Fe2+) and energy that can be obtained from the spore reserves. This stage was blocked by agents that inhibit ATP formation but not by antibiotics that inhibit macromolecular synthesis. The second stage, swelling, needed an exogenous carbon source and was not blocked by mitomycin C. In this stage, the spores exhibited the highest cytochrome oxidase and catalase activities and respiratory quotient. The last stage, germ tube emergence, required additional carbon and nitrogen sources. Ammonium compounds were superior to nitrate. Dry weight remained constant during the stages of darkening and swelling, with a rapid increase from the moment of germ tube emergence. Optimum pH and temperature for germination were 8.0 and 45 degrees C, respectively. Heat treatment (55 degrees C for 10 min) had no effect on germination. The fine structure of the spore underwent important changes during germination. The wall of the swollen spore became stratified and the inner layer was continuous with the germ tube wall. Macromolecular synthesis occurred in the sequence RNA, protein and then DNA. Rifampicin, streptomycin and mitomycin C prevented synthesis when added at the start of incubation. The same effect was obtained if the addition was made during germination, except with mitomycin C which inhibited DNA, but not RNA and protein synthesis.     

Effects of snout dimensions on the hydrodynamics of suction feeding in juvenile and adult seahorses

Seahorses give birth to juveniles having a fully functional feeding apparatus, and juvenile feeding behaviour shows striking similarities to that of adults. However, a significant allometric growth of the snout is observed during which the snout shape changes from relatively short and broad in juveniles to relatively long and slender in adults. Since the shape of the buccal cavity is a critical determinant of the suction performance, this snout allometry will inevitably affect the suction feeding ability. To test whether the snout is optimised for suction feeding throughout an ontogeny, we simulated the expansion of different snout shapes varying from extremely long and slender to short and broad for juvenile and adult snout sizes, using computational fluid dynamic models. Our results showed that the snout diameter at the start of the simulations is involved in a trade-off between the realizable suction volume and expansion time on the one hand (improving with larger initial diameters), and maximal flow velocity on the other hand (improving with smaller initial diameters). Moreover suction performance (suction volume as well as maximal attainable flow velocity) increased with decreasing snout length. However, an increase in snout length decreases the time to reach the prey by the cranial rotation, which may explain the prevalence of long snouts among syngnathid fishes despite the reduced suction performance. Thus, the design of the seahorse snout revolves around a trade-off between the ability to generate high-volume suction versus minimisation of the time needed to reach the prey by the cranial rotation.     

水淹对枫杨幼苗光合生理特征的影响

通过设置对照(CK)、连续性水淹(CF)和间歇性水淹(PF)3个水分处理,模拟三峡库区库岸带土壤水分变化,研究乡土树种枫杨当年实生幼苗的生理生态适应机制.结果表明:不同水分处理均显著影响枫杨幼苗的光合作用、生物量积累和生长.与CK相比,CF和PF组枫杨幼苗除胞间CO2浓度升高,净光合速率(Pn)、气孔导度(gs)均显著降低.其变化趋势是枫杨幼苗的Pn、gs在试验初期下降,然后逐渐恢复或趋于稳定.随着处理时间的延长,CF和PF组枫杨幼苗的总生物量、根生物量、茎生物量、叶生物量、株高和地径均呈现上升趋势.CF和PF组的总生物量、根生物量、叶生物量和株高,以及PF组的茎生物量均显著低于CK,而CF组的茎生物量与CK无显著差异,其地径还高于CK.枫杨幼苗具有耐受水湿而不耐水淹-干旱交替的生理生态特征.