企业在燃料和农业领域的合作推动生物技术的实用化

企业间的合作在医药行业习以为常。但是在利用生物技术的燃料、农业领域．以商业化为目的的合作才陆续开始。在医药品方面打头阵的生物技术的实用化．在其他领域正在加速进行。     

1987年大兴安岭特大火灾后北坡森林景观生态恢复评价

通过空间直观景观模型(LAND IS)来探讨大兴安岭北坡1987年特大森林火灾后,在目前这种恢复方案下以及完全依靠天然更新下森林景观的长期动态,通过对比研究来评价目前所采取的恢复措施是否能够有效地恢复森林资源。研究结果表明:1987年大火后所采取的恢复措施可以在很大程度上增加针叶树种在该区所占的比例,而相应地降低阔叶树种的比重。由于在目前的恢复措施下,不仅大面积地种植针叶树种,同时也采伐了大量的成、过熟林,使落叶松和樟子松的蓄积量在开始阶段不仅没有增加,反而有大幅度的下降。但最终,所更新的幼苗逐渐成材,落叶松和樟子松的蓄积量逐渐超过天然更新下的。在演替前期,白桦的蓄积量在这2种模拟方案下相差不大;而在演替后期,白桦在天然更新方案下的蓄积量要高于在目前恢复措施下的蓄积量。另外,物种在不同的区域的变化动态又有较大的差异。在重度火烧区,由于人为种植大量的针叶树种,使它们所占的面积比例明显高于完全依靠天然更新下其在重度火烧区所占的比例;而阔叶树种则相反,在完全依靠天然更新方案下所占的面积百分比明显高于在目前恢复方案下的比例。而在未火烧和轻中度火烧区,由于火后人为采伐大量的落叶松和樟子松,使这2个树种在该区所占的比例在开始100多年里要低于完全依靠天然更新下的比例,随后逐渐高于天然更新下所占的比例,但相差不大;阔叶树种则正好相反。另外,目前恢复方案不仅极大地改变了主要物种在该区所占的比例,而且还明显的影响了它们的空间分布格局。     

对哺乳动物细胞中重组DNA产物安全性的探讨

人们期待着在不久的将来,用重组哺乳动物细胞的表达系统来提供药用产品,某些产品现已在临床试验。 尽管哺乳动物细胞培养昂贵是其缺陷。但已证明它在表达重组DNA的药用产品方面极为有用,在微生物表达系统,这些产品的质量常有问题。除了在开始需进行基因操作外,重组细胞的培养过程在概念上与传统的产品所用的生产方法是一样的。现有的科学工具已能确证从哺乳动物细胞连续培养的株系中分离的重组产物,在用于人类医疗方面是安全的。     

稻-麦轮作系统土壤水解酶及氧化还原酶活性对开放式空气CO2浓度增高的响应

研究了FACE条件下(CO2浓度增加200μmol·mol^-1)水稻、小麦不同生育期0～10cm土层土壤脲酶、磷酸酶、芳基硫酸酯酶、脱氢酶活性的变化．结果表明,FACE条件下,土壤脲酶活性在冬小麦生育前期低于对照,在孕穗期高于对照;在水稻生育前期高于对照,在成熟期低于对照．磷酸单酯酶活性在冬小麦生育期高于对照;在水稻分蘖期高于对照,在生育后期(拔节期、抽穗期和成熟期)低于对照．芳基硫酸酯酶活性在小麦越冬期和孕穗期低于对照,在分蘖期和成熟期高于对照;在水稻生育期间均高于对照．脱氢酶活性在小麦和水稻的生育前期低于对照,在后期高于对照．     

低亲和性神经营养素受体p75在人胚胎视网膜中的表达

p75神经营养素受体在视网膜的发育以及再生过程中发挥着重要的作用,而在人类视网膜中的分布状况尚未被研究. 利用免疫组织化学方法,在光镜水平下确定了p75在人胚胎发育5、6和7个月的视网膜中的分布情况. 在视网膜神经节细胞层出现最强的p75免疫阳性反应,在其他各层也有较弱的免疫阳性反应. 在胚胎6、7月的视网膜中,主要由Müller细胞的终足构成的内界膜上出现了比较强的p75表达. p75在人胚胎视网膜中的分布情况与大鼠视网膜中很类似,主要表达在Müller细胞, 在神经节细胞上也可能有表达.     

生长休止蛋白7在成年大鼠肾脏、心脏和肝脏中的差异表达

目的研究生长休止蛋白7（Gas7）在成年大鼠肾脏、心脏和肝脏的表达。方法成年SD大鼠16只,分别采用逆转录聚合酶链反应（RT-PCR）方法和免疫组织化学方法检测Gas7基因mRNA和蛋白在成年SD大鼠肾脏、心脏和肝脏的表达,并进行图像分析和统计学处理。结果RT—PCR结果显示,Gas7mRNA在肾脏高表达,在心脏的表达弱于肾脏（P〈0．05）,而在肝脏的表达最弱,基本检测不到。免疫组化结果显示,在肾脏中,Gas7免疫阳性产物在近髓肾单位的近曲小管呈强阳性反应,在集合管表达较弱,在肾小球和其余肾小管未见表达;在心脏中,Gas7免疫阳性产物均匀分布于心肌细胞,呈中等强度反应,弱于肾脏（P〈O．05）;在肝脏中,Gas7蛋白未见明显表达,与其mRNA在肝脏的表达相似。结论Gas7在大鼠肾脏、心脏和肝脏表达的不同,尤其在肾脏组织分布的差异性,提示Gas7在成年大鼠肾脏和心脏结构以及功能的维持中可能起着重要作用。     

GUS基因在梨转化植株中的稳定表达

以获得的梨转GUS基因植株的试管苗为试材,转基因植株的叶片在含卡那霉素的再生培养基上可成功再生不定梢,这些不定梢在含卡那霉素的生根培养基上可诱导生根。再生不定梢经组织化学检测仍表现出GUS基因的表达。而非转基因植株的叶片在含卡那霉素的再生培养基上不能产生不定梢,显示外源基因在转基因植株中的表达在无性繁殖后代中是稳定遗传的。     

寄主挥发物、叶色和表皮毛在美洲斑潜蝇寄主选择中的作用

在室内条件下 ,初步研究了寄主挥发物、叶色和表皮毛在美洲斑潜蝇寄主选择中的作用。在嗅觉仪试验中 ,寄主叶片挥发物对美洲斑潜蝇雌成虫没有明显的引诱作用 ;在叶色反应试验中 ,美洲斑潜蝇雌成虫在叶子圆片上停留的时间明显大于在滤纸圆片上停留的时间 (p<0 .0 1) ,其在有叶片区域分布的数量明显多于空白对照 (p <0 .0 1) ;在表皮毛试验中 ,美洲斑潜蝇在无毛叶片上的产卵量明显大于在有毛叶片上的产卵量 (p <0 .0 1)。上述结果表明 ,在对寄主的定向和定位过程中 ,美洲斑潜蝇的视觉起着重要的作用 ,而嗅觉不起作用 ;叶片表皮毛有抑制产卵的作用     

Paxillin在大鼠胰腺发育不同时期表达的研究

目的:研究桩蛋白(Paxillin,Pxn)在胰腺发育不同阶段的表达和细胞定位.方法:运用RT-PCR技术检测Pxn在大鼠胰腺发育不同阶段的mRNA表达水平;运用免疫组织化学检测不同时期桩蛋白在胰腺的定位.结果:RT-PCR结果显示Pxn的mRNA表达量胚胎期高于新生和成年期;免疫组织化学结果显示在不同发育时期桩蛋白不仅在外分泌胰腺有表达,而且在胰岛也有表达.结论:具有调节细胞聚集、粘附迁移功能的桩蛋白可能参与出生后胰岛重塑.     

在水生与气生状态下石莼光合作用对光照和温度的响应

如何对付由于高潮时的水生状态与低潮时的气生状态高频率循环所导致的不同环境条件,是潮间带海藻的光合作用所面临的独特问题。对采自汕头沿岸的石莼(Ulva lactuca)在水生和气生不同状态下光合作用对光照和温度的响应特性进行了测定,以探讨这种常见的潮间带绿藻在潮汐循环背景下的光合特性。在气生状态下,光饱和净光合速率(Pmax)随气生暴露时间的变化模式可以很好地用三次方程进行描述,而温度影响方程的系数;当水分损失为15％时,石莼的Pmax增加至最大值,然后Pmax随进一步脱水而下降,在水分损失为80％时下降至0。温度对Pmax的影响在水生状态下比在气生状态下更大。气生状态下（充分水化）Pmax在10℃时显著小于水生状态下的值,而在30℃时则相反。在10℃时,气生干出时间在6 h 以内,或在20℃时,气生干出时间在2.2 h 以内,石莼的净碳固定量在气生状态下比在水生状态下要大;而在30℃时,在气生状态下的净碳固定量比总是小于在水生状态下的净碳固定量。认为石莼在低潮气生状态下与在高潮水生状态下光合特性及净碳固定存在差异,但这种差异与环境温度及叶状体的水分状态有关。     

Effect of zinc,phosphorus and nitrogen on tryptophan concentration in rice grains grown on limed and unlimed soils

Summary The effects of Zn, P, N and CaCO₃ on tryptophan concentration in rice grain were studied in greenhouse at Haryana Agricultural University. Zinc application upto 20 ppm increased tryptophan concentration in rice grain. Zn-EDTA gave highest increase followed by ZnSO₄ and then ZnO. Liming at the rate of 4 and 8 per cent decreased tryptophan concentration significantly. Phosphorus application upto 100 ppm also decreased tryptophan significantly but Zn in combination with P increased tryptophan and overcame negative effect of P. Nitrogen application upto 120 ppm increased tryptophan concentration. There was positive interaction between Zn and N. Ammonium sulphate gave highest tryptophan followed by ammonium nitrate and then urea. The tryptophan concentration ranged between 766 ppm and 2011 ppm in paddy grain. The lowest tryptophan concentration was in the plants treated with 8 per cent lime in absence of added Zn and highest with 10 ppm Zn through Zn-EDTA. Department of Soils.     

Tryptophan catabolism by tryptophan pyrrolase in rat liver. The effect of tryptophan loads and changes in tryptophan pyrrolase activity.

We investigated how changes in tryptophan pyrrolase activity and tryptophan loads affect the breakdown of tryptophan was estimated by injecting rats with [ring-2-14-C]tryptophan and measuring respiratory 14-CO2. We concluded, contrary to previous reports, that induction of tryptophan pyrrolase definitely will increase the rate of tryptophan breakdown. Tryptophan loads also increase tryptophan breakdown even in circumstances where there is no increase in tryptophan pyrrolase activity, presumably by increasing the saturation of the enzyme. After a tryptophan load (50 mg per kg) the increase in liver tryptophan concentration lasts only 30 min. The rapid return of liver tryptophan to normal may be due partly to the high turnover rate of liver tryptophan. We estimate that tryptophan pyrrolase degrades tryptophan in vivo at a rate that is equivalent to the whole liver tryptophan concentration in 7.5 min or less.     

Tryptophan and serotonin metabolism after sustained tryptophan infusion

In an attempt to elucidate the effects of sustained administration of tryptophan on serotonin synthesis and turnover in mammalian brain, mini-osmotic pumps containing tryptophan or vehicle were implanted in albino mice for 24 and 96 h. Despite the extremely low dose of tryptophan administered by these pumps (8–12 mg/kg-day) statistically significant treatment effects were apparent with both treatment durations. Plasma and brain tryptophan concentrations varied in unison, and were inversely related to the tryptophan degradative capabilities of the liver as reflected in tryptophan pyrrolase activity. After 24 h of tryptophan infusion the hepatic enzyme activity was elevated and tryptophan values were no different from controls, and after 96 h the hepatic enzyme activity was reduced and tryptophan values in treated animals were greater than controls. Serotonin was elevated in treated animals after 24 h, but not after 96 h despite the elevated tryptophan concentration at this time. The turnover of serotonin, as evidenced by 5-hydroxyindoleacetic acid concentrations, was not significantly affected by either treatment.Hepatic degradation of tryptophan thus seemed to be an important determinant of total plasma tryptophan, and brain tryptophan values paralleled plasma tryptophan. It appears that serotonin biosynthesis is regulated by factors other than tryptophan availability when the latter is chronically elevated.     

Leucine and tryptophan metabolism in rats.

The rate of tryptophan metabolism in isolated liver cells from animals fed on a high-leucine diet was greater than for cells from control animals. Leucine inhibited tryptophan metabolism and tryptophan uptake in isolated liver cells, probably by competing for membrane transport. Leucine had no effect on tryptophan 2,3-dioxygenase in vitro. 4-Methyl-2-oxovalerate increased tryptophan oxidation in incubations containing albumin, by displacing bound tryptophan and increasing the availability of the amino acid to the cell. The results suggest that, under extreme conditions, when the availability of tryptophan is low, leucine may be pellagragenic.     

Tryptophan properties in fluorescence and functional stability of plasminogen activator inhibitor 1

Plasminogen activator inhibitor 1 harbors four tryptophan residues at positions 86, 139, 175, and 262. To investigate the contribution of each tryptophan residue to the total fluorescence and to reveal the mutual interactions of the tryptophan residues and interactions with the other amino acids, 15 mutants in which tryptophan residues have been replaced by phenylalanines were constructed, purified, and characterized. Conformational distribution analysis revealed that the tryptophan mutants have a similar conformational distribution pattern as wild-type plasminogen activator inhibitor 1. Mutants in which tryptophan residue 175 was replaced by a phenylalanine displayed an increased functional half-life of the active conformation, whereas the functional half-life of mutants in which tryptophan residue 262 was replaced by a phenylalanine was substantially decreased. Comparative analysis of the fluorescence lifetimes, the extinction coefficients, and the quantum yields of the individual tryptophan residues demonstrates that tryptophan residue 262 gives the highest contribution to the total fluorescence. The other tryptophan residues have a very low quantum yield. In the wild-type protein, the fluorescence of all tryptophan residues is partially quenched as compared to the mutants that contain single tryptophan residues, due to conformational effects. The fluorescence of tryptophan residue 262 is very likely also partially quenched by energy transfer to tryptophan residue 175.     

Studies on tryptophan residues of Abrus agglutinin. Stopped-flow kinetics of modification and fluorescence-quenching studies.

The presence of two essential tryptophan residues/molecule was implicated in the binding site of Abrus agglutinin [Patanjali, Swamy, Anantharam, Khan & Surolia (1984) Biochem. J. 217, 773-781]. A detailed study of the stopped-flow kinetics of the oxidation of tryptophan residues revealed three classes of tryptophan residues in the native protein. A discrete reorganization of tryptophan residues revealed three classes of tryptophan residues in the native protein. A discrete reorganization of tryptophan residues into two phases was observed upon ligand binding. The heterogeneity of tryptophan exposure was substantiated by quenching studies with acrylamide, succinimide and Cs+. Our study revealed the microenvironment of tryptophan residues to be hydrophobic, and also the presence of acidic amino acid residues in the vicinity of surface-localized tryptophan residues.     

Intralipid and free plasmatic tryptophan in vitro

In an attempt to investigate the role of the lipidic emulsion Intralipid in the development of metabolic encephalopathy in a patient showing high free tryptophan levels, the relationship between lipidic emulsion and free tryptophan was examined in in vitro experiments. The addition of intralipid to normal serum produces an immediate increase in non-esterified fatty acids and a parallel rise in free tryptophan. Moreover, when serum with intralipid is incubated at 37 degrees C, the lipases release new non-esterified fatty acids and the free tryptophan increases proportionally. The non-esterified fatty acid content of intralipid was found to be 12 +/- 2 mEq X 1(-1). An inverse correlation was seen between free tryptophan and different serum albumin concentrations. It is concluded that intralipid causes an increase in free tryptophan levels. It is known that in vivo free tryptophan modulates 5-hydroxytryptamine synthesis and thus may be considered a possible causal agent for encephalopathy.     

Indolic Substances in Plasma, Cerebrospinal Fluid, and Frontal Cortex of Human Subjects Infused with Saline or Tryptophan

Psychiatric patients undergoing the psychosurgical operation of stereotactic subcaudate tractotomy were infused intravenously with either saline or L-tryptophan (15 mg/kg/h). Plasma, lumbar cerebrospinal fluid (CSF), ventricular CSF and a specimen of frontal cortex were collected. The relationships of plasma concentrations of substances claimed to influence brain tryptophan concentration (total tryptophan, free tryptophan, large neutral amino acids) with the concentration of tryptophan in the cortex and CSF were investigated. Tryptophan infusion resulted in plasma tryptophan values comparable to those found after oral doses used in treating depression or insomnia, and about sixfold increases of tryptophan in the cerebral cortex. Increased brain 5-hydroxytryptamine synthesis was indicated by significant rises of CSF 5-hydroxyindoleacetic acid. The concentration of plasma free tryptophan was a better predictor than plasma total tryptophan of cortex tryptophan concentration. As all correlation coefficients of plasma versus brain or plasma versus ventricular CSF tryptophan concentrations were decreased when allowance was made for differences of concentration of large neutral amino acids, the results suggest that the role of these substances within their physiological range as inhibitors of tryptophan transport to the brain may previously have been overemphasised.     

Effects of insulin and streptozotocin-induced diabetes on brain tryptophan and serotonin metabolism in rats

Previous work by other authors has shown hat insulin administration increases brain tryptophan levels and serotonin (5–HT) metabolism. The present study partially replicates these results and tests whether these effects could be due to insulin-induced hypoglycemic stress, since stressers such as immobilization or food deprivation also increase brain tryptophan and 5-HT metabolism. Ingestion of a dextrose solution by rats administered insulin (2 I.U./kg) prevents the extreme fall in blood glucose concentration and rise in plasma corticosterone following insulin injections alone. This treatment, however, produces a larger increase in brain tryptophan (30%) than insulin-injected rats allowed only tap water. The greater accumulation of brain tryptophan may reflect an additive effect of the endogenously released insulin to that exogenously administered, since ingestion of the dextrose solution could trigger insulin secretion. In addition, brain tryptophan and 5-HT metabolism were measured in streptozotocin-diabetic rats maintained on several different feeding schedules to control for the effects of hyperphagia. All groups of diabetics showed significant decreases of approx 30% in brain tryptophan concentrations, while 5-HT metabolism was unchanged. This deficit in brain tryptophan is reversed within 2 h after insulin administration (2 I.U./kg). These results indicate that changes in brain tryptophan and 5-HT metabolism following insulin injections are not due to hypoglycemic stress, and that brain tryptophan is low in diabetics but increases above normal after administration of insulin. The results are discussed with respect to the effects of insulin on plasma levels of the neutral amino acids and a possible direct effect of insulin on the uptake of tryptophan by brain.     

Guinea-pig liver tryptophan pyrrolase. Absence of detectable apoenzyme activity and of hormonal induction by cortisol and possible regulation by tryptophan

1. When assayed in fresh homogenates, guinea-pig liver tryptophan pyrrolase exists only as holoenzyme. It does not respond to agents that activate or inhibit the rat liver enzyme in vitro. Only by aging (for 30min at 5 degrees C) does the guinea-pig enzyme develop a requirement for ascorbate. 2. The guinea-pig liver enzyme is activated by the administration of tryptophan but not cortisol, salicylate, ethanol or 5-aminolaevulinate. 3. The tryptophan enhancement of the guinea-pig liver pyrrolase activity is prevented by 0, 34 and 86% by pretreatment with actinomycin D, cycloheximide or allopurinol respectively. 4. The guinea-pig liver tryptophan pyrrolase is more sensitive to tryptophan administration than is the rat enzyme. On the other hand, the concentrations of tryptophan in sera and livers of guinea pigs are 45-52% less than those in rats. 5. It is suggested that tryptophan may regulate the activity of guinea-pig liver tryptophan pyrrolase by mobilizing a latent form of the enzyme whose primary function is the detoxication of its substrate.