Enhancement of In Vitro Growth of Pathogenic Bacteria by Norepinephrine: Importance of Inoculum Density and Role of Transferrin

Norepinephrine is a stress hormone that enhances bacterial growth. We examined the effects of a small inoculum on the norepinephrine-induced growth of species previously reported to be unaffected by norepinephrine. The results indicated that a reduced inoculum density is essential for observing norepinephrine-induced effects. Additional studies using serum-free media suggested that transferrin plays a role in norepinephrine-induced growth.     

Propagule pressure and persistence in experimental populations

Average inoculum size and number of introductions are known to have positive effects on population persistence. However, whether these factors affect persistence independently or interact is unknown. We conducted a two-factor experiment in which 112 populations of parthenogenetic Daphnia magna were maintained for 41 days to study effects of inoculum size and introduction frequency on: (i) population growth, (ii) population persistence and (iii) time-to-extinction. We found that the interaction of inoculum size and introduction frequency-the immigration rate-affected all three dependent variables, while population growth was additionally affected by introduction frequency. We conclude that for this system the most important aspect of propagule pressure is immigration rate, with relatively minor additional effects of introduction frequency and negligible effects of inoculum size.     

Controls for rhizosphere microorganisms to study effects of vesicular-arbuscular mycorrhizae on Artemisia tridentata

Seven treatments were set up to test the effects of vesicular-arbuscular (VA) mycorrhizal fungi and other rhizosphere microorganisms on the growth of Artemisia tridentata ssp. tridentata. Soil sievings had no significant effect on root or shoot mass. Spores and surface-sterile spores were a poor inoculum source, but roots and fresh soil caused 45–75% mycorrhizal infection. Whereas root-inoculated plants still had low growth responses by the end of the experiment, fresh soil inoculum caused the greatest response, and partial fresh inoculum caused a lesser response. These results suggest that fresh soil is an appropriate inoculum for this plant-fungal-soil system, and that the major effect on plant growth of the fresh soil inoculum is from the mycorrhizal fungi and not from the other microorganisms, because the sievings had no effect on plant growth. In addition, soil dilution plating of saprophytic fungi showed 85% species similarity between sterile and fresh soil inoculum by the end of the experiment. Since the effects of non-VA microorganisms are complex and varied, we suggest that researchers work out the type of mycorrhizal controls that best suit their system.     

The effect of glucagon on vasoconstriction and vascular escape from nerve- and norepinephrine-induced constriction of the hepatic artery of the cat

Glucagon, in the anesthetized cat, was capable of dilating the hepatic artery to the same extent and in a dose-dependent manner when administered directly into the hepatic artery or into the portal vein. Portal venous infusions of glucagon did not inhibit nerve- or norepinephrine-induced vasoconstriction of the hepatic artery in contrast to previous reports in the dog. Rather, at certain doses, glucagon mildly potentiated the vasoconstriction induced by both constrictor stimuli. Vascular escape from nerve- and norepinephrine-induced constrictor responses was found to be inhibited by glucagon in a dose-dependent manner. Glucagon infusion is the first intervention reported to modulate vascular escape in the hepatic artery. Owing to its similar effects on nerve- and exogenous norepinephrine-induced responses, glucagon appears to be acting at a postsynaptic site. Therefore, we suggest that in the cat, glucagon is not an inhibitory modulator of nerve- and norepinephrine-induced vasoconstriction, but rather may potentiate the constrictor response in a postsynaptic manner.     

Effects of pyruvate on the growth of normal and transformed hamster embryo fibroblasts

The growth of NIL and NILpy hamster embryo fibroblasts was determined in the presence and absence of pyruvate as a component of the growth medium. It was demonstrated that NIL cells respond to the presence of pyruvate by decreasing cell doubling time, glucose utilization, glutamine utilization, and increasing lactate production with the effects being more pronounced at low inoculum densities. Polyoma-virus transformed NIL cells (NILpy) demonstrate none of the above effects upon pyruvate addition regardless of initial cell inoculum density.     

装液量和接种量对红发夫酵母生长和虾青素积累的影响

在摇瓶中研究了不同接种量和装液量对红发夫酵母PhaffiarhodozymaAs2 .15 5 7生长及虾青素积累的影响。结果表明在 5 0 0ml摇瓶中装液量为 5 0ml时对红发夫酵母生长及虾青素合成有利 ,接种量为 10 %有利于菌体生长和虾青素合成。     

Calreticulin inhibits the MEK1,2-ERK1,2 pathway in alpha 1-adrenergic receptor/Gh-stimulated hypertrophy of neonatal rat cardiomyocytes

In cardiac myocytes, stimulation of alpha(1)-adrenoceptor (AR) leads to a hypertrophic phenotype. The G(h) protein (transglutaminase II, TGII) is tissue type transglutaminase and transmits the alpha(1B)-adrenoceptor signal with GTPase activity. Recently, it has been shown that the calreticulin (CRT) down-regulates both GTP binding and transglutaminase activities of TGII. To elucidate whether G(h) mediates norepinephrine-stimulated intracellular signal transductions leading to activation of extracellular signal-regulated kinases (ERKs) and neonatal rat cardiomyocyte hypertrophy, we examined the effects of G(h) on the activation of ERKs and inhibitory effects of CRT on alpha(1)-adrenoceptor/G(h) signaling. In neonatal rat cardiomyocytes, norepinephrine-induced ERKs activation was inhibited by an alpha(1)-adrenoceptor blocker (prazosin), but not by an beta-adrenoceptor blocker (propranolol). Overexpression of the G(h) protein stimulated norepinephrine-induced ERKs activation, which was inhibited by alpha-adrenoceptor blocker (prazosin). Co-overexpression of G(h) and CRT abolished norepinephrine-induced ERKs activation. Taken together, norepinephrine induces hypertrophy in neonatal rat cardiomyocytes through alpha(1)-AR stimulation and G(h) is partly involved in norepinephrine-induced MEK1,2/ERKs activation. Activation of G(h)-mediated MEK1,2/ERKs was completely inhibited by CRT.     

Relationship of seed source and arbuscular mycorrhizal fungi inoculum type to growth and colonization of big bluestem (Andropogon gerardii)

Big bluestem (Andropogon gerardii) is a key grass of tallgrass prairies and is commonly included in restoration projects. In many cases, it has been found to benefit significantly from arbuscular mycorrhizal (AM) fungi, however results have varied under non-sterile soil conditions. This study investigated the effects of two types of AM fungi inoculum (commercial and prairie) on growth and root colonization of big bluestem from five different seed sources grown in non-sterile soils. Seed sources were collected from five remnant prairies in the Tallgrass Prairie Peninsula located in the Midwestern United States. Growth of big bluestem and root length colonized by AM fungi was highly variable among seed source treatments. Overall, percentage of root length colonized by AM fungi was positively correlated with the total dry weight of plants, and plants that received inoculum generally grew better than those that did not receive inoculum. Inoculum treatment affected both big bluestem growth and percentage root length colonized and there was an interaction between seed source and inoculum treatment relative to colonization. Root colonization responses were not significantly different between the prairie and commercial inoculum types, although there was a significant response between plants that received additional inoculum as opposed to no additional inoculum. Seed sources from Ohio and Illinois had the highest biomasses and greatest percentage of root length colonized while plants from Wisconsin and Missouri grew poorly and had low root colonization. These results demonstrate the importance of considering both seed source and inoculum type before the incorporation of AM inoculum to prairie restoration projects.     

Production of a Laccase and Decrease of the Phenolic Content in Canola Meal during the Growth of the Fungus Pleurotus ostreatus in Solid State Fermentation Processes

Solid state fermentation of canola meal was carried out with the fungus Pleurotus ostreatus DAOM 197961, which is a producer of laccase. The aim of this study was to examine the effects of moisture content, inoculum size, homogenisation of inoculum and particle size of canola meal on the growth of the fungus, the production of a laccase and the decrease of the content of sinapic acid esters (SAE) in a solid state process. The results showed that the optimum moisture content, which was varied in the media between 50% and 75%, for the growth and enzyme production was 60%. The initial rate of SAE content decrease was faster in the media with 70% and 75% moisture than in those with lower moisture levels. In the study of the effects of inoculum concentration in the range of 1.1 mg to 5.5 mg/g of the medium, it was found that larger amounts of biomass and enzyme were produced in the media with inoculum concentrations from 1.1 mg to 3.3 mg/g of the medium than in the media with a higher inoculum concentration. The final and approximately the same concentrations of SAE were reached at the same time regardless of the inoculum concentration. Considering that the fungus formed pellets under the conditions at which it was grown during the inoculum preparation, it was necessary to break them by homogenisation prior to their utilisation as an inoculum. The homogenisation was carried out during a period between 15s and 200s. Although higher biomass concentrations and enzyme activities were obtained in the media which were inoculated with the inoculum homogenised for 15s and 30s, the maximum enzyme activities and biomass concentrations were reached in the media inoculated with the inoculum, which was homogenised for 120s and 200s. The time of inoculum homogenisation did not influence the kinetics of the SAE decrease. When the effects of the particle size of canola meal on the process were studied, it was found that larger particles of the meal in the solid media were more favourable for the production of the biomass and enzyme, and for a faster decrease of the SAE content than those of smaller sizes. From the obtained results it can be concluded that the tested variables have a significant influence on the growth of the fungus Pleurotus ostreatus DAOM 197961, the production of laccase and the decrease of the SAE content in canola meal. The data could be useful for the development of a solid state process for the production of laccase and for the decrease of the phenolics content in canola meal.     

Regulation of mammalian cell growth by autocrine growth factors: analysis of consequences for inoculum cell density effects

Effects of inoculum cell density on mammalian cell growth in culture have been observed in a variety of experimental systems. Although these effects have been attributed generally to medium conditioning by the cells, there has previously been no quantitative theory proposed for this phenomenon based on developments in molecular and cell biology. In this article, we offer such a theory founded on the regulatory action of autocrine growth factors. A particularly relevant example of these is platelet- derived growth factor (PDGF), which is produced by fibroblastic cells in response to stimulation by transforming growth factor beta (TGFbeta), a common serum constituent, and provides a mitogenic signal for the same cells. A simple mathematical model for the production, diffusive transport, and binding of autocrine growth factors to cell surface receptors, coupled to a model for the dependence of cell proliferation on growth factor receptor binding allows prediction of initial cell population growth rate as a function of inoculum cell density. We focus on situations involving anchorage-dependent cell growth, in which the cells are attached to a surface. A number of clear results are obtained, most notably the following: 1) for cells cultured on spherical microcarrier bead surfaces, the inoculum cell density needed to produce a given growth rate is linearly proportional to the bead radius; and 2) all other factors being equal, the inoculum cell density on a unit surface area basis needed to produce a given growth rate is greater for spherical microcarrier surfaces than for flat culture dish surfaces. These two results are consistent with the experimental observations of Hu and coworkers(1,2) for fibroblast growth in minimal medium plus serum. The model also allows elucidation of the influence of other system parameters, both biological and physical, on initial cell proliferation rate and the inoculum cell density dependence.     

Vitex glabrata R.Br.悬浮培养细胞的生长条件优化及20-羟基蜕皮激素的生产

研究了培养基、植物生长调节素以及接种量对Vitex glabrata R.Br.悬浮培养细胞的生长情况以及对20-羟基蜕皮激素形成的作用。当细胞在添加有2.0mg/LBAP(6-苯甲酸嘌呤)和1.0mg/L2,4-D的Gamborg’s B5培养基中培养时细胞生长和20-羟基蜕皮激素的形成达到了最高水平。当接种量为20%PCV(积压细胞体积)时观察到了20-羟基蜕皮激素的最高产量,大约是1.1mg/(L.d)。实验数据也表明当接种量增加到20%PCV时,产量提高了7倍。     

The Influence of Temperature on Meloidogyne incognita on Soybean

The effects of temperature and initial inoculum density of Meloidogyne incognita on soybean growth and nematode reproduction were investigated in greenhouse temperature tanks and in controlled-growth chambers. The interactions of initial inoculum density (P_i) and soil temperature in effects on shoot growth were adequately described by multiple-regression models. At the highest temperatures (30 or 32/28 C), moderate to high inoculum killed many plants. A P_i of 27,000 eggs/15-cm-diam pot retarded shoot growth at 26 C. Only the greatest P_i (81,000 eggs/15-cm pot) suppressed shoot growth at 18, 22, or 20/16 C. Inoculation with 3,000 or 9,000 eggs/plant resulted in heavier root systems at all temperatures except 30 C. At that temperature, 9,000 eggs suppressed root growth. At 18 and 26 C, a Pi of 81,000 eggs was required to retard root growth. Nematode reproduction was related directly to temperature and P_i except at a density of 81,000 eggs/15-cm pot.     

The effect of inoculum age and solid versus liquid propagation on inoculum quality of an industrial Bacillus licheniformis strain

Shorter lag phases were obtained in cultivations of Bacillus licheniformis using early-compared to late-stationary growth phase inocula and using liquid versus solid propagation medium. Flow cytometry and fluorescence ratio imaging microscopy (FRIM) after staining with 5(6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE), confirmed that liquid early-stationary growth phase inoculum had a higher vitality and was more homogeneous than solid late-stationary growth phase inoculum. DNA-microarray analyses indicated that liquid early-stationary growth phase inoculum was in a more active state in terms of cell multiplication whereas solid late-stationary growth phase inoculum was induced to some spore formation potentially causing delayed growth initiation.     

Pro-apoptotic versus anti-apoptotic properties of dietary resveratrol on tumoral and normal cardiac cells

Resveratrol is a natural dietary polyphenol found in grape skin, red wine, and various other food products. Resveratrol has proved to be an effective chemopreventive agent for different malignant tumors. It has also been shown to prevent vascular alterations such as atherosclerosis and inflammatory-associated events. In view of these observations, we investigated the anti-proliferative and pro-apoptotic activities of resveratrol on a tumoral cardiac cell line (HL-1 NB) derived from mouse tumoral atrial cardiac myocytes. These effects were compared with those found on normal neonatal mouse cardiomyocytes. HL-1 NB cells and neonatal cardiomyocytes were treated with resveratrol (5, 30, and/or 100 μM) for different times of culture (24, 48, and/or 72 h). Resveratrol effects were determined by various microscopical and flow cytometric methods. After resveratrol treatment, a strong inhibition of tumoral cardiac HL1-NB cell growth associated with a loss of cell adhesion was observed. This cell proliferation arrest was associated with an apoptotic process revealed by an increased percentage of cells with fragmented and/or condensed nuclei (characteristic of apoptotic cells) identified after staining with Hoechst 33342 and by the presence of cells in subG1. At the opposite, on normal cardiomyocytes, no cytotoxic effects of resveratrol were observed, and a protective effect of resveratrol against norepinephrine-induced apoptosis was found on normal cardiomyocytes. Altogether, the present data demonstrate that resveratrol (1) induces apoptosis of tumoral cardiac HL1-NB cells, (2) does not induce cell death on normal cardiomyocytes, and (3) prevents norepinephrine-induced apoptosis on normal cardiomyocytes.     

灵芝多糖发酵工艺条件的研究

通过对灵芝菌As5.504发酵,研究不同糖源、氮源、生长因子及接种量、装液量对灵芝细胞生长和灵芝多糖生产的影响。结果表明当以葡萄糖,蛋白胨,VB5分别为碳源、氮源、生长因子及接种量、装液量分别为7.5mL、150mL时,为As5.504产多糖最佳发酵工艺条件,产多糖量最高为0.940g·L-1。     

Adrenomedullin modulates hemodynamic and cardiac effects of angiotensin II in conscious rats

We examined whether adrenomedullin, a vasoactive peptide expressed in the heart, modulates the increase in blood pressure, changes in systolic and diastolic function, and left ventricular hypertrophy produced by long-term administration of ANG II or norepinephrine in rats. Subcutaneous administration of adrenomedullin (1.5 microg.kg(-1).h(-1)) for 1 wk inhibited the ANG II-induced (33.3 microg.kg(-1).h(-1) sc) increase in mean arterial pressure by 67% (P < 0.001) but had no effect of norepinephrine-induced (300 microg.kg(-1).h(-1) sc) hypertension. Adrenomedullin enhanced the ANG II-induced improvement in systolic function, resulting in a further 9% increase (P < 0.01) in the left ventricular ejection fraction and 19% increase (P < 0.05) in the left ventricular fractional shortening measured by echocardiography, meanwhile norepinephrine-induced changes in systolic function were remained unaffected. Adrenomedullin had no effect on ANG II- or norepinephrine-induced left ventricular hypertrophy or expression of hypertrophy-associated genes, including contractile protein and natriuretic peptide genes. The present study shows that adrenomedullin selectively suppressed the increase in blood pressure and augmented the improvement of systolic function induced by ANG II. Because adrenomedullin had no effects on ANG II- and norepinephrine-induced left ventricular hypertrophy, circulating adrenomedullin appears to act mainly as a regulator of vascular tone and cardiac function.     

Modelling and Estimation of the Relative Potential for Infection of Winter Wheat by Inoculum of Gaeumannomyces graminis Derived from Propagules and Infected Roots

Abstract A model was developed to estimate the mean number of infections of seminal roots of wheat exposed to two sources of inoculum of the take-all fungus Gaeumannomyces graminis var. tritici , in an experimental system. The sources comprise discrete propagules of initial, soil inoculum and infected roots of volunteer plants that had been infected by the initial inoculum, prior to the growth of crop plants. The model was based on the probability of escape from infection by individual roots ofthe crop plants. Parameter estimation was by maximum likelihood. A model was first fitted to data for infection of roots from the soil inoculum. This yielded estimates for the efficiency of soil inoculum to cause infection in the absence of volunteer plants. The parameter for efficiency of infection by soil inoculum was resolved into components for inoculum density, survival of inoculum and the probability of success of individual propagules. The model was extended to include simultaneous exposure of crop roots to soil inoculum and to root inoculum on the volunteer plants. The presence of volunteer seedlings prior to sowing of crop plants resulted in an increase in the effectiveness of inoculum to cause disease. Sowing date and soil condition, as affected by addition of sand, were shown to have significant effects on the efficiency of both sources of inoculum.     

Arbuscular mycorrhizal fungi enhance spotted knapweed growth across a riparian chronosequence

Arbuscular mycorrhizal fungi (AMF) mediate nutrient uptake that accelerates plant growth and reproduction. Thus, AMF may promote plant invasions often observed along rivers. We assessed the importance of AMF in improving growth of the invasive species, spotted knapweed (Centaurea stoebe), during succession of riparian vegetation along a flood plain in Montana, USA. We grew spotted knapweed with and without AMF in soils collected from riparian sites ranging from 1 to 72 years old and measured the plant’s growth response to AMF. We observed variability in relative effects of AMF, with greatest growth benefits in recently deposited alluvial sediments. We then separated effects of soil and inoculum source by growing spotted knapweed with soils and inocula collected from young or old sites and found that growth responses were greatest in young soils regardless of inoculum source. Our results demonstrate that AMF directly benefit growth of spotted knapweed, especially in soils that typify early successional sites on this alluvial flood plain.     

搅拌式生物反应器悬浮培养水母雪莲细胞的研究

应用 2L通气搅拌式生物反应器一步批式培养水母雪莲细胞。采用倾斜式搅拌桨代替透平桨 ,研究了搅拌转速、通气量和接种量对细胞生长和黄酮合成的影响 ,发现在 75r min、70 0～1000L min和 4.0～ 5.0gDCW L接种量下细胞生长和黄酮合成比较好。经过 12d培养细胞干重达 13.8gDCW L ,黄酮产量 416mg L ,黄酮含量占细胞干重的 30%。水母雪莲细胞生长及黄酮合成的进程表明 ,黄酮积累与细胞生长呈正相关。对细胞聚集体分布的研究发现 ,流变压力使细胞聚集体分裂 ,使反应器中细胞生长受到影响 ,黄酮产量较摇瓶中降低     

Relationship between the quality of the inoculum material and carminomycin biosynthesis by a culture of Actinomadura carminata]

The effect of the inoculum mycelium quality on carminomycin biosynthesis by Actinomadura carminata was studied. The time of the organism growth on the culture medium containing cornsteep liquor continued for 6 hours without losing by the inoculum of its seeding qualities during that period. The mycelium growth in the inoculum was more intensive under conditions of moderate aeration, i.e. 0.98-2.64 mg O2H1-min. Anincrease in the aeration rate up to 18.56 mg O2/1-min resulted in the growth suppression up to 40 per cent. No correlation between the aeration rate during the inoculum growth and the culture capacity for carminomycin biosynthesis and of the content of the complex in active components the fermentation medium were observed, when a 5-10 per cent of inoculum was used.