Molecular cloning and expression of the gene for a major leucine-rich protein from human hepatoblastoma cells (HepG2)

Summary The human hepatoblastoma cell line, HepG2, exhibits an array of stable properties in culture that have made it a popular cell culture model for studies on regulation of liver-specific gene expression and properties of hepatoma cells. In contrast to other hepatoma cell lines, HepG2 cells overexpress a characteristic detergent-extractable, wheat germ lectin-binding protein with apparent molecular mass of 130 kDa. Using an antibody to screen a phage expression library of HepG2 complementary DNA (cDNA), we identified and cloned a 4734 base pair cDNA which codes for a 130-kDa leucine-rich protein (lrp130) when expressed in transfected cells. The deduced sequence of lrp130 exhibits sequences weakly homologous to the consensus sequence for the ATP binding site in ATP-dependent kinases and the protein kinase C phosphorylation site of the epidermal growth factor receptor. Consistent with the higher levels of expression of lrp130 antigen, Northern hybridization analysis indicated that HepG2 cells express high levels of the major 4.8 kilobase lrp130 mRNA relative to other hepatoma cells. Although currently of unknown function, lrp130 may be of utility as a marker for liver cell lineages represented by the HepG2 cell line.     

Substantial overproduction of antibodies by applying osmotic pressure and sodium butyrate

Much of the current cell technology has enabled increased antibody production levels due to judicious nutrient feeding to raise cell densities and design better bioreactors. This study demonstrates that hybridomas can be hyperstimulated to produce higher immunoglobulin (lg) levels by suppressing cell growth and increasing culture longevity through adaptation to higher osmolarity media and addition of sodium butyrate. Prior to adaptation, cells placed in higher osmotic pressures (350 and 400 mOsm) were severely suppressed in growth down to 25% of the control (300 mOsm), although total lg titers achieved were similar to the control, approximately 140 mg/L. After a week of adaptation to 350 and 400 mOsm media, cell growth was not as dramatically suppressed, but considerably higher lg levels were attained at these elevated osmolarities. The highest yield of 265 mg/L was obtained at 350 mOsm compared to 140 mg/L at 300 mOsm, while maximum viable cell numbers dropped from 35 x 10(5) cells/mL to 31 x 10(5) cells/mL and culture longevity was extended by 20 h more than the control. Sodium butyrate, known to enhance protein production in other cell types, was then supplemented at a range of concentrations between 0.01 and 0.4 mM to the 350 mOsm culture to further enhance the lg levels. Butyrate at a concentration of 0.1 mM, in combination with osmotic pressure at 350 mOsm, further elevated the lg levels to 350 mg/L. Concomitantly, maximum viable cell numbers were reduced to 22 x 10(5) cells/mL, but culture longevity was extended by 40 h in the 0.1 mM butyrate supplemented culture compared to the control condition. Specific antibody productivity, q(Mab), continued to stay high during the stationary phase and was further elevated during the decline phase: thus, overall lg levels can be increased by 2.3 times by combining osmotic pressure and butyrate treatment. (c) 1993 John Wiley & Sons, Inc.     

OxyR介导的细菌氧化胁迫应答调控机制研究进展

OxyR属于LysR型转录因子家族的氧化胁迫调控蛋白,是细菌抵抗氧化胁迫压力的重要调控因子。OxyR能够通过调控过氧化氢酶和过氧化物酶等抗氧化基因的表达清除H₂O₂、参与铁代谢控制胞内过氧化物的产生以及修复生物大分子氧化损伤,从而抵抗氧化胁迫。OxyR的基因表达调控功能依赖于其还原态和氧化态之间的转变,改变调控蛋白对下游基因调控区的亲和能力。氧化态OxyR识别启动子区的结合序列,激活或抑制过氧化氢酶等基因的表达。还原态和氧化态的转换依赖于在氧化状态下分子间二硫键的形成。本文综述了近年来细菌OxyR调控基因表达的最新研究进展,有助于深入理解OxyR在细菌抵抗氧化胁迫的作用方式,为相关致病菌的防治奠定分子基础。     

珠三角城市群区域生态系统服务供需均衡关系

生态系统服务供需均衡关系分析为生态系统管理提供了详实的科学基础信息。基于土地利用、气象和社会经济等多源数据,采用遥感反演、水量平衡方程、修正的通用土壤流失方程和植被净生产力模型(CASA)等方法,分别评估了1990—2015年珠三角城市群粮食供给、产水服务、固碳释氧和土壤保持服务;采用相关社会经济指标核算了生态系统服务的需求量;进而揭示了栅格、县域和市域多尺度的生态系统服务供需均衡关系及其时空演变特征。研究发现：(1)除粮食供给外,产水服务、固碳释氧和土壤保持服务呈波动性增加的趋势,增幅分别为52.2%、21.8%和73.4%;在空间上表现为中部平原地区服务水平低,低山丘陵地带高的分布特征。(2)除土壤保持服务外,粮食供给、产水服务和固碳释氧服务需求量不断增加,增幅分别为10.1%、17.5%和769.4%;中部平原地区生态系统服务需求量大,低山丘陵地带小。(3)除土壤保持和产水服务外,粮食供给、固碳释氧及综合服务供需指数不断下降,其中,2015年粮食供给和固碳释氧供需指数分别为-0.47和-0.71。研究结果可为决策者了解区域的生态系统服务供需均衡匹配状况提供科学基础信息,直接服务并...     

滇南地区桃树种植模式对土壤有机碳组分及碳库管理指数的影响

经果林种植可改变土壤质量、改善生态环境,同时具有较高的经济效益。合理的种植模式可通过物种间的互补性提高资源利用效率,改善土壤碳库质量并提高综合效益。为探讨桃树种植模式对土壤有机碳组分及碳库管理指数的影响,以云南省开远市不同桃树种植模式(桃树单种-SP和桃树南瓜套种-PP)为研究对象,以毗邻的天然林地(CK)为对照,分析不同种植模式下活性碳库,即高锰酸钾氧化有机碳(POXC)、颗粒有机碳(POC),惰性有机碳库即矿物结合态有机碳(MAOC)在0—40 cm土层的分布情况,明确土壤有机碳组分与土壤理化性质的关系;计算碳库活度指数(CPAI)、碳库指数(CPI)以及碳库管理指数(CPMI),明确不同桃树种植模式的碳库变化情况。结果表明：桃树种植模式和对照的土壤有机碳组分的含量均随着土层深度的增加而减少,平均土壤有机碳(SOC)含量为：14.68 g/kg(CK)>9.57 g/kg(PP)>8.58 g/kg(SP)。平均活性有机碳组分所占比例与POC/MAOC均表现为：SP>CK>PP,PP的活性有机碳比例较低,具有较高的有机碳稳定性。两种桃树种植模式的CPMI在1...     

Nutrient uptake relationship to root characteristics of rice

Data on root parameters and distribution are important for an improved understanding of the factors influencing nutrient uptake by a crop. Therefore, a study was conducted on a Crowley silt loam at the Rice Research and Extension Center near Stuttgart, Arkansas to measure root growth and N, P and K uptake by three rice (Oryza sativa L.) cultivars at active tillering (36 days after emergence (DAE)), maximum tillering (41 DAE), 1.25 cm internode elongation (55 DAE), booting (77 DAE) and heading (88 DAE). Soil-root core samples were taken to a depth of 40 cm after plant samples were removed, sectioned into 5 cm intervals, roots were washed from soil and root lengths, dry weights and radii were measured. Root parameters were significantly affected by the soil depth × growth stage interaction. In addition, only root radius was affected by cultivar. At the 0- to 5-cm soil depth, root length density ranged from 38 to 93 cm cm^-3 throughout the growing season and decreased with depth to about 2 cm cm^-3 in the 35- to 40-cm depth increment. The increase in root length measured with each succeeding growth stage in each soil horizon also resulted in increased root surface area, hence providing more exposed area for nutrient uptake. About 90% of the total root length was found in the 0- to 20-cm soil depth throughout the season. Average root radius measured in the 0- to 5-cm and 35- to 40-cm depth increments ranged from 0.012 to 0.013 cm and 0.004 to 0.005 cm, respectively throughout the season. Total nutrient uptake by rice differed among cultivars only during vegetative growth. Differences in total nutrient uptake among the cultivars in the field appear to be related to absorption kinetics of the cultivars measured in a growth chamber study. Published with permission of the Arkansas Agricultural Experiment Station.     

The Drosophila Meiotic Recombination Gene Mei-9 Encodes a Homologue of the Yeast Excision Repair Protein Rad1

Meiotic recombination and DNA repair are mediated by overlapping sets of genes. In the yeast Saccharomyces cerevisiae, many genes required to repair DNA double-strand breaks are also required for meiotic recombination. In contrast, mutations in genes required for nucleotide excision repair (NER) have no detectable effects on meiotic recombination in S. cerevisiae. The Drosophila melanogaster mei-9 gene is unique among known recombination genes in that it is required for both meiotic recombination and NER. We have analyzed the mei-9 gene at the molecular level and found that it encodes a homologue of the S. cerevisiae excision repair protein Rad1, the probable homologue of mammalian XPF/ERCC4. Hence, the predominant process of meiotic recombination in Drosophila proceeds through a pathway that is at least partially distinct from that of S. cerevisiae, in that it requires an NER protein. The biochemical properties of the Rad1 protein allow us to explain the observation that mei-9 mutants suppress reciprocal exchange without suppressing the frequency of gene conversion.     

Failure of short term stimulation to reduce sarcoplasmic reticulum Ca2+-ATPase function in homogenates of rat gastrocnemius

To examine the effect of short term intense activity on sarcoplasmic reticulum (SR) Ca²⁺ sequestering function, the gastrocnemius (G) muscles of 11 anaesthetized male rats (weight, 411±8 g,X±SE) were activated using supramaximal, intermittent stimulation (one train of 0.2 msec impulses per sec of 100 msec at 100 Hz). Homogenates were obtained from stimulated white (WG-S) and red (RG-S) tissues, assayed for Ca²⁺ uptake and maximal Ca²⁺ ATPase activity and compared to contralateral controls (WG-C, RG-C). Calcium uptake (nmoles/mg protein/min) determined using Indo-l and at [Ca²⁺]_f concentrations between 300–400 nM was unaffected (p>0.05) by activity in both WG (6.14+0.43 vs 5.37+0.43) and RG (3.21+0.18 vs 3.07+0.20). Similarly, no effect (p>0.05) of contractile activity was found for maximal Ca²⁺ ATPase activity (mole/mg protein/min) determined spectrophotometrically in RG (0.276+0.03 vs 0.278+0.02). In WG, Ca²⁺ ATPase activity was 15% higher in WG-S compared to WG-C (0.412+0.03 vs 0.385+0.04). Repetitive stimulation resulted in a reduction in tetanic tension of 74% (p<0.05) by 2 min in the G muscle. By the end of the stimulation period, ATP concentration was reduced (p<0.05) by 57% in the WG and by 47% in the RG. These results indicate that the repeated generation of maximal tetanic force, at least for short term periods, need not adversely affectin vitro homogenate determination of Ca²⁺ sequestering function in spite of severe alterations in energy potential and that some other mechanism must be involved to explain the depression in Ca²⁺ uptake and Ca²⁺ ATPase activity previously noted with short term intense exercise.     

甘蔗细平象的研究

甘蔗细平象Trockorhopalus humeralis Chevrolat是云南甘蔗上的一种毁灭性地下害虫,以幼虫和成虫在地下蔗头内为害,为害期8-10个月。据1989年调查,受害蔗每亩损失500-3000kg,严重的无收。此虫1年发生1代,以成虫在蔗头内越冬,有喜湿性,不能飞翔,主要通过沟河流水传播。在河川坝地,沙壤土中虫口较多;宿根年限越长的甘蔗受害越重。建议蔗稻轮作;缩短甘蔗宿根年限;早春翻挖有虫蔗蔸烧毁;结合新植蔗下种,宿根蔗松蔸培土施用甲基异柳磷或铁灭克等颗粒杀虫剂进行防治。