991 resultados para culture condition
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The acute toxic effect of the pesticide cypermethrin to Daphnia magna HB was examined. D. magna HB was exposed to cypermethrin at concentrations of 0, 1, 3, 5, 7, and 9 mg/L for 24 In. Data showed that the 24 h-LC50 of cypermthrin on D. magna HB was 4.81 mg/L. In contrast, the 24 h-LC50 of K2Cr2O7 (the national standard toxicant) to Daphnia magna was 0.38 mg/L in the current study. Results indicated that the Daphnia magna was very sensitive to pesticides. In addition, the effects of the culture condition(such as hardness, temperature and DO etc.) on Daphnia magna HB was also studied.
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本文根据我们实验室建立的发酵产物中辅酶Q10定性定量检测方法,筛选得到一株可以代谢产生较多辅酶Q10的野生菌株放射形土壤杆菌(Agrobacterium radiobacter No.50)。 为了提高放射形土壤杆菌的辅酶Q10的产量,本实验利用液体培养研究了单因素对菌株辅酶Q10产量的影响,并用正交法确定了最佳液态发酵条件。最佳发酵培养基是:葡萄糖20g,蔗糖40g, 硫酸铵10g,玉米浆30g, 酵母膏3g,K2HPO4 3g,MgSO4.7H2O 1g,蒸馏水1000mL,pH 7.0-7.2。最佳发酵条件是:转接斜面菌种到种子培养基, 转速220r/min、温度28。C培养24h后,转入发酵培养基(250mL三角拼装液量为50mL,pH 7.0), 接种量为10%,转速220r/min、温度28。C,培养120h。在此条件下,菌体湿重约为50g/L,辅酶Q10含量约为20mg/L。 本文以放射形土壤杆菌为出发菌株进行诱变育种,以期获得辅酶Q10高产菌。根据微生物育种原理、参照辅酶Q10的代谢调控机制,以野生型放射形土壤杆菌(Agrobacterium radiobacter No.50)为出发菌株,采用紫外线和亚硝基胍复合诱变技术,依次筛选得到菌体提取物M抗性菌ARM-7、烟草提取物T抗性菌株ARMT-26、Vk3抗性菌株ARMTV-25、链霉素抗性菌株ARMTVS-32,菌株ARMTVS-32产量达到了36.8mg/L,与原始出发菌株相比,产量提高了77%。 研究了茄尼醇、对羟基苯甲酸、橘子皮提取物D、胡萝卜提取物E、烟草提取物对ARMTVS-32合成辅酶Q10的影响,结果表明这些物质对菌体合成辅酶Q10有一定促进作用,添加0.2g/L茄尼醇时,辅酶Q10含量提高了17%,达到了40.7mg/L;添加1.2g/L橘子皮提取物D时,辅酶Q10含量提高了13.8%,达到了39.6mg/L;添加0.5g/L胡萝卜提取物E时,辅酶Q10含量提高了25.3% ,达到了43.6mg/L;添加8g/L烟草提取物时,辅酶Q10含量提高了12.6%,达到了39.2mg/L。 Production of Coenzyme- Q10 (CoQ10) by fermentation is considered as a process with broad prospects.Quantitative Analysis of CoQ10 in the culture of microbe by TLC—UV spectrophotometry was developed, by using this method we got the strain Agrobacterium radiobacter,which was isolated from forest soil of southwest of China. The effect of the single factor on CoQ10-production ability of the strain was examined by liquid cultured, and its best optimum cultivation conditions were established by orthogonal method. The results showed that the optimum fermentation conditions were as following: carbon sources glucose 20g/L,sucrose 40g/L; nitrongen sources (NH4)2SO4 10g/L,maize liquid 30g/L;yeast extract 3g; K2HPO4 3g/L,MgSO4.7H2O 1g/L; initial pH was 7 and volume of medium(medium volume vs flask volume) was 50mL/500mL, incubating for 120h on a rotary shaker at 220 rpm and 28℃.Under these conditions, the biomass and CoQ10 concentration reached 50g/L and 20mg/L respectively. According to the biosynthesis mechanism of CoQ10 and breeding theory, CoQ10 over-production strains were screened by UV--NTG. mutation using Agrobacterium radiobacter No.50 as parent strain. A microbe-juice resistant mutant ARMTVS-32, which also could resist tobacco-juice, VK3 and streptomycin, was screened out from an agar plate. The CoQ10 content of ARMTVS-32 reached 36.8mg/L, which was 77% higher than the initial strain. In addition, We discussed the effects of some organic substrates on the synthesis of CoQ10 in ARMTVS-32. The results showed that solanesol, orange juice D, carrot juice E and tobacco juice could promote the CoQ10 accumulation in the cells. The CoQ10 content of ARMTVS-32 reached 40.7mg/L when added 0.2g/L solanesol,it reached 39.6mg/L when added 1.2g/L orange juice D,it reached 43.6mg/L when added 0.5g/L carrot juice E. it reached 39.2mg/L when added 8g/L tobacco juice.
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本论文以红薯淀粉的双酶法水解液为碳源,从19 株红色酵母中筛选出一株油脂含量较高的菌株掷孢酵母(Sporobolomyces reseus)As.2.618。为了提高掷孢酵母(S.reseus)As.2.618 的油脂产量,考察了培养基组成对该菌生长情况及油脂积累的影响。用均匀设计法对培养基组成进行了优化,由DPS软件得出的优化结果为:还原糖103g/L、酵母粉11.5g/L、磷酸二氢钾0.3g/L、硫酸镁0.15g/L。生物量可达19.23 g/L,油脂含量为3.875 g/L。研究了添加二价离子对该菌的生长及油脂积累的影响,结果表明Zn2+对该菌生长和油脂积累都有显著促进作用。研究了发酵条件以及添加氧载体正十二烷对该菌发酵的影响,表明添加正十二烷有利用于该菌生长与油脂积累。得出最佳发酵条件是:在还原糖103g/L、酵母粉11.5g/L、磷酸二氢钾0.3g/L、硫酸镁0.15g/L。添加30mg/L 硫酸锌,接种量为5%,在24h 后添加2g/L 的碳酸钙和2%(v/v)正十二烷,pH6.0 培养温度为27℃,转速为200r/min,培养时间为7 天的条件下,该菌生物量干重可达35.05g/L,油脂含量也达11.98g/L。Lipid is one of the basic material for life-sustaining activities andimportant industrial materials. As lipid resources mainly come from the animal andthe plant, the problem of lipid lack is encountered at times. The lipid frommicroorganisms is the substitute and superior to the above lipid with a short period ofproduction and much cheaper fermentation materials such as agricultural and sidelineproducts or wastes of crop.Thus large scale production and broad application ofmicrobial lipid will be efficient not only in substitute of the animal and the plant lipidfor food and industrial field , but also inducing a new way leading to solve the energyproblem.For the purpose of exploring the characteristics of lipid production of redyeasts from sweet potato starch hydrolysates. 19 red yeasts are screened for thecapability of lipid producing and one strain Sporobolomyces reseus As.2.618 withsuperior performance is sellected.To improve the Sporobolomyces reseus As.2.618’s capability of lipidaccumulation , the components of the medium, which may influence the growth of thestrain and the lipid yield have been studied. To get the optimum mediumcomponents ,the “uniform design” was used .The DPS software gave the optimummedium component is: reducing sugar 103 g/L、yeast extract 11.5 g/L、KH2PO4 0.3g/L、MgSO4 0.15 g/L. The biomass could reach up to 19.23 g/L and lipid yield 3.87g/L with the above composition of fermentation medium.Furthermore the fermentation conditions , addition of the divalent metal ionsand the oxygen vector to increase the strain’s lipid producing capability are tested.The optimum condition is : reducing sugar 103 g/L、yeast extract 11.5 g/L、KH2PO40.3 g/L、MgSO4 0.15 g/L,Adding 30mg/L ZnSO4,and adding 2g/L CaCO3 2%(v/v)n-dodecane after 24h’s fermentation. the optimal fermentation condition were asfollow :30ml medium in the 500ml flask with initial pH 6.0,the flasks with 5%inoculation volume were at 200r/min shaking speed for 7d’s fermentation at27 .Under this kind of condition the high biom ¡æ ass which reach to 35.05 g/L could begot ,the yield of lipid also could reach to 11.98g/L.
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Eutrophication is becoming a serious problem in coastal waters in many parts of the world. It induces the phytoplankton blooms including 'Red Tides', followed by heavy economic losses to extensive aquaculture area. Some cultivated seaweeds have very high productivity and could absorb large quantities of N, P, CO2, produce large amount of O-2 and have excellent effect on decreasing eutrophication. The author believes that seaweed cultivation in large scale should be a good solution to the eutrophication problem in coastal waters. To put this idea into practice, four conditions should be fulfilled: (a) Large-scale cultivation could be conducted within the region experiencing eutrophication. (b) Fundamental scientific and technological problems for cultivation should have been solved. (c) Cultivation should not impose any harmful ecological effects. (d) Cultivation must be economically feasible and profitable. In northern China, large-scale cultivation of Laminaria japonica Aresch. has been encouraged for years to balance the negative effects from scallop cultivation. Preliminary research in recent years has shown that Gracilaria lemaneiformis (Bory) Daws. and Porphyra haitanensis Chang et Zheng are the two best candidates for this purpose along the Chinese southeast to southern coast from Fujian to Guangdong, Guangxi and Hong Kong. Gracilaria tenuistipitata var. liui Chang et Xia is promising for use in pond culture condition with shrimps and fish.
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Gracilaria lemaneiformis (Bory) Daws has been extensively cultivated as a source of commercial agar and the ecomaterials in Shenao Bay, Guangdong Province, Jiaozhou Bay, Shandong Province and other waters in China. This paper examines the in situ suspended farming of G. lemaneiformis using raft cultivation under different conditions and its effects on nutrient removal in the laboratory. The results showed that cultivated Gracilaria grew well in both Shenao Bay and Jiaozhou Bay. The biomass of Gracilaria increased from 50 to 775 g m(-1) (fresh weight) during 28 days, with special growth rate (SPG) 13.9% d(-1) under horizontal cultivation in Jiaozhou Bay. Light, temperature, nutrient supply, as well as cultivation treatments such as initial density, and depth of suspension seaweed were important to the growth of Gracilaria. The highest biomass production was observed in the horizontal culture condition (0.0 m) and 0.5-1.5 m deep layer in Jiaozhou Bay. However, the highest growth rate in Shenao Bay appeared under the lowest initial stocking density treatment. In the laboratory, the aquarium experiments (fish and seaweed culture systems) demonstrated that Gracilaria was able to remove inorganic nutrients effectively. The concentration of NH4+-N decreased by 85.53% and 69.45%, and the concentration of PO4-P decreased 65.97% and 26.74% in aquaria with Gracilaria after 23 days and 40 days, respectively. The results indicate that Gracilaria has the potential to remove excess nutrient from coastal areas, and the large-scale cultivation of G. lemaneiformis could be effective to control eutrophication in Chinese coastal waters. (c) 2005 Elsevier B.V. All rights reserved.
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Evidence that persistent environmental pollutants may target the male reproductive system is increasing. The male reproductive system is regulated by secretion of testosterone by testicular Leydig cells, and perturbation of Leydig cell function may have ultimate consequences. 3-Methylsulfonyl-DDE (3-MeSO2-DDE) is a potent adrenal toxicants formed from the persistent insecticide DDT. Although studies have revealed the endocrine disruptive effect of 3-MeSO2-DDE, the underlying mechanisms at cellular level in steroidogenic Leydig cells remains to be established. The current study addresses the effect of 3-MeSO2-DDE on viability, hormone production and proteome response of primary neonatal porcine Leydig cells. The AlamarBlue™ assay was used to evaluate cell viability. Solid phase radioimmunoassay was used to measure concentration of hormones produced by both unstimulated and Luteinizing hormone (LH)-stimulated Leydig cells following 48h exposure. Protein samples from Leydig cells exposed to a non-cytotoxic concentration of 3-MeSO2-DDE (10μM) were subjected to nano-LC-MS/MS and analyzed on a Q Exactive mass spectrometer and quantified using label-free quantitative algorithm. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) were carried out for functional annotation and identification of protein interaction networks. 3-MeSO2-DDE regulated Leydig cell steroidogenesis differentially depending on cell culture condition. Whereas its effect on testosterone secretion at basal condition was stimulatory, the effect on LH-stimulated cells was inhibitory. From triplicate experiments, a total of 6804 proteins were identified in which the abundance of 86 proteins in unstimulated Leydig cells and 145 proteins in LH-stimulated Leydig cells was found to be significantly regulated in response to 3-MeSO2-DDE exposure. These proteins not only are the first reported in relation to 3-MeSO2-DDE exposure, but also display small number of proteins shared between culture conditions, suggesting the action of 3-MeSO2-DDE on several targeted pathways, including mitochondrial dysfunction, oxidative phosphorylation, EIF2-signaling, and glutathione-mediated detoxification. Further identification and characterization of these proteins and pathways may build our understanding to the molecular basis of 3-MeSO2-DDE induced endocrine disruption in Leydig cells.
A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária
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DUARTE, E. N. ; CARVALHO, Andréa Vasconcelos ; PINHEIRO, E. G. ; CARVALHO, Luciana Moreira ; NOGUEIRA, N. M. F. . A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária. Informação & Sociedade. Estudos, João Pessoa - PB, v. 10, n. 2, p. 190-210, 2000
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To evaluate the nutritional value of palm kernel meal on the performance Nile tilapia (Oreochromis niloticus), five isonitrogenous (30% crude protein), isoenergetic (2800 kcal/kg of digestible energy), and isofibrous (10% crude fiber) diets, with increasing levels of palm kernel meal (0, 7, 14, 21, 28 and 35%). were ad libitum fed to fingerlings with initial body weight of 1.52g +/- 0.04, housed per 120 days in 35 aquarium of 60 liters with fingerlings each. The environmental culture condition were monitored during all experimental period. Statistical analyses of recorded data were performed through polynomial regression models. To determine the apparent digestibility, macroscopic analysis of viscera and microscopic analysis of intestine epithelium were performed. The obtained results indicated that the inclusion of palm kernel meal up to 35% Nile tilapia diets did not affects the apparent digestibility and did not induce any pathological effects on viscera and intestinal epithelium.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Hypoxia-inducible factor-1 alpha (HIF-1α) plays a critical role in survival and is associated with poor prognosis in solid tumors. The role of HIF-1α in multiple myeloma is not completely known. In the present study, we explored the effect of EZN2968, an locked nucleic acid antisense oligonucleotide against HIF-1α, as a molecular target in MM. A panel of MM cell lines and primary samples from MM patients were cultured in vitro in the presence of EZN2968 . Under normoxia culture condition, HIF-1α mRNA and protein expression was detectable in all MM cell lines and in CD138+ cells from newly diagnosed MM patients samples. Significant up-regulation of HIF-1α protein expression was observed after incubation with IL6 or IGF-I, confirming that HIF-1α can be further induced by biological stimuli. EZN2968 efficiently induces a selective and stable down-modulation of HIF-1α and decreased the secretion of VEGF released by MM cell. Treatment with EZN2968 gave rise to a progressive accumulation of cells in the S and subG0 phase. The analysis of p21, a cyclin-dependent kinase inhibitors controlling cell cycle check point, shows upregulation of protein levels. These results suggest that HIF-1α inhibition is sufficient for cell cycle arrest in normoxia, and for inducing an apoptotic pathways.. In the presence of bone marrow microenvironment, HIF-1α inhibition blocks MAPK kinase pathway and secretion of pro-surviaval cytokines ( IL6,VEGF,IL8) In this study we provide evidence that HIF-1α, even in the absence of hypoxia signal, is expressed in MM plasma cells and further inducible by bone marrow milieu stimuli; moreover its inhibition is sufficient to induce a permanent cell cycle arrest. Our data support the hypothesis that HIF-1α inhibition may suppress tumor growth by preventing proliferation of plasma cells through p21 activation and blocking pro-survival stimuli from bone marrow microenvironment.
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The aim of this thesis was to investigate the regenerative potential of alternative sources of stem cells, derived from human dental pulp (hDPSCs) and amniotic fluid (hAFSCs) and, specifically, to evaluate their capability to be committed towards osteogenic and myogenic lineages, for the eventual applicability of these stem cells to translational strategies in regenerative medicine of bone and skeletal muscle tissues. The in vitro bone production by stem cells may represent a radical breakthrough in the treatment of pathologies and traumas characterized by critical bone mass defects, with no medical or surgical solution. Human DPSCs and AFSCs were seeded and pre-differentiated on different scaffolds to test their capability to subsequently reach the osteogenic differentiation in vivo, in order to recover critical size bone defects. Fibroin scaffold resulted to be the best scaffold promoting mature bone formation and defect correction when combined to both hDPSCs and hAFSCs. This study also described a culture condition that might allow human DPSCs to be used for human cell therapy in compliance with good manufacturing practices (GMPs): the use of human serum (HS) promoted the expansion and the osteogenic differentiation of hDPSCs in vitro and, furthermore, allowed pre-differentiated hDPSCs to regenerate critical size bone defects in vivo. This thesis also showed that hDPSCs and hAFSCs can be differentiated towards the myogenic lineage in vitro, either when co-cultured with murine myoblasts and when differentiated alone after DNA demethylation treatment. Interestingly, when injected into dystrophic muscles of SCID/mdx mice - animal model of Duchenne Muscular Dystrophy (DMD) - hDPSCs and hAFSCs pre-differentiated after demethylating treatment were able to regenerate the skeletal muscle tissue and, particularly, to restore dystrophin expression. These observations suggest that human DPSCs and AFSCs might be eventually applied to translational strategies, in order to enhance the repair of injured skeletal muscles in DMD patients.
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IκB kinase α (IKKα) is one kinase subunit of the IKK complex that is responsible for NF-κB activation. Previous studies have shown that IKKα determines mouse keratinocyte terminal differentiation independent of the NF-κB pathway. Accumulating evidence suggests that IKKα functions as a tumor suppressor in skin carcinogenesis; however, the downstream pathways mediating this function are largely unknown. By using primary cultured keratinocytes, we found that Ikkα-/- cells developed aneuploidy and underwent spontaneous immortalization and transformation while wild type cells underwent terminal differentiation in the same culture condition. Using proteomic analysis we identified nucleophosmin (NPM), a centrosome duplication regulator, as an IKKα substrate. We further demonstrated that IKKα interacted with NPM and colocalized with NPM on the centrosome, suggesting that NPM is a physiological substrate of IKKα. Loss of IKKα reduced centrosome-bound NPM and promoted abnormal centrosome amplification, which contributed to aneuploidy development. Detailed analysis revealed that ablation of IKKα target site serine-125 of NPM induced destabilization of NPM hexamers, disrupted NPM association with centrosomes, and resulted in abnormal centrosome amplification. Re-introduction of IKKα rescued the defect in Ikkα-/- keratinocytes. Thus, IKKα is required for maintaining proper centrosome duplication by phosphorylating NPM. ^ UV is the major etiological agent for human skin cancer and UV-induced mouse skin carcinogenesis is one of the most relevant experimental models for human skin carcinogenesis. Thus, we further evaluated IKKα function in UV-induced skin carcinogenesis in Ikkα+/- mice. We demonstrated that IKKα is also critical in UV skin carcinogenesis, as evidenced by increased tumor multiplicity and reduced tumor latency in Ikkα+/- mice after chronic UVB treatment. Reduced expression of IKKα decreased UV-induced apoptosis and promoted accumulation of P53 mutations in the epidermis. This indicates that IKKα is critical for UV-induced apoptosis in vivo and thus prevents mutation accumulation that is important for tumor development. ^ Together, these findings uncover previously unknown in vivo functions of IKKα in centrosome duplication and apoptosis, thus providing a possible mechanism of how loss of IKKα may contribute to skin carcinogenesis. ^
A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária
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DUARTE, E. N. ; CARVALHO, Andréa Vasconcelos ; PINHEIRO, E. G. ; CARVALHO, Luciana Moreira ; NOGUEIRA, N. M. F. . A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária. Informação & Sociedade. Estudos, João Pessoa - PB, v. 10, n. 2, p. 190-210, 2000
A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária
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DUARTE, E. N. ; CARVALHO, Andréa Vasconcelos ; PINHEIRO, E. G. ; CARVALHO, Luciana Moreira ; NOGUEIRA, N. M. F. . A cultura organizacional influenciando o comportamento do capital humano na biblioteca universitária. Informação & Sociedade. Estudos, João Pessoa - PB, v. 10, n. 2, p. 190-210, 2000
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O crescimento celular da microalga Haematococcus pluvialis e a bioprodução de carotenoides são influenciados pelas diferentes condições de cultivo. Dentre os corantes naturais, a astaxantina tem importante aplicação farmacêutica, cosmética e na indústria de alimentos. Este pigmento além de colorir, possui propriedades biológicas, dentre elas a atividade antioxidante. A produção de astaxantina através do cultivo de H. pluvialis pode alcançar até 4% do peso seco da microalga. O objetivo desse trabalho foi avaliar o crescimento celular, bem como a produção de carotenoides pela microalga Haematococcus pluvialis em diferentes condições de cultivo e a atividade antioxidante dos extratos carotenogênicos. Foram utilizados os meios autotróficos Blue Green-11 (BG-11), BAR (Barbera Medium) e BBM (Bold Basal Medium) e os meios mixotróficos BBM e glicose e BBM e acetato de sódio, empregando 10 ou 20% de inóculo em pHs iniciais de 6, 7 ou 8, aeração de 0,30 L.min-1 , sob iluminância de 6 Klx, 24±1ºC durante 15 dias em fotobiorreatores de 1 L. A concentração celular foi avaliada diariamente através de leitura de absorvância a 560 nm. A ruptura celular foi realizada através de 0,05 g de células secas com 2 mL de dimetilsulfóxido e a concentração de carotenoides totais determinada a partir de leitura espectrofotométrica a 474 nm. Os meios de cultivo BG-11, BBM e glicose e BBM e acetato de sódio apresentaram, respectivamente, o maior crescimento celular e produção de carotenoides totais de 0,64, 1,18 e 0,68 g.L-1 , e 3026,66, 2623,12 e 2635,38 µg.g-1 , empregando 10% de inóculo em pH inicial de 7. Com base nesses resultados, foram selecionados esses três meios para dar continuidade ao trabalho. O meio de cultivo BBM e acetato de sódio obteve o melhor valor de concentração celular máxima, com 1,29±0,07 g.L-1 e de carotenoides totais 5653,56 µg.g-1 empregando pH inicial de 7 e concentração de inóculo de 20%. Este meio foi selecionado para a realização dos cultivos com injeção de 30 % de CO2 uma vez ao dia durante 1 hora, realizados durante 22 dias, em pH inicial de 7 e 20% de inóculo, com 30% de injeção de CO2 uma vez ao dia durante 1 hora. Nestas condições o crescimento celular alcançou o máximo de 1,13 g.L-1 (10 dias), carotenoides totais específicos de 2949,91 µg.g-1 e volumétricos de 764,79 µg.g-1 .L-1 (22 dias). A capacidade antioxidante dos extratos carotenogênicos também foi avaliada pelos métodos DPPH, FRAP e ABTS, não sendo possível quantificá-la através do DPPH e FRAP. Por outro lado, utilizando o método ABTS, em 90 minutos de reação, o poder de inibição encontrado foi de 35,70 % μg-1 . Assim, a condição que mais se destaca é a utilização do meio de cultivo BBM e acetato de sódio, com pH inicial 7, com 20% de inóculo, 0,30 L.min-1 de aeração, 6 Klx e 24±1ºC, uma vez que o crescimento celular e a bioprodução de carotenoides foi significativamente superior quando comparada às demais condições estudadas. Além disso, os carotenoides produzidos pela H. pluvialis, nesta condição, apresentaram capacidade antioxidativa.