Nitrate depletion in the riparian zone of a small woodland stream

Field enrichments with nitrate in two spring-fed drainage lines within the riparian zone of a small woodland stream near Toronto, Ontario showed an absence of nitrate depletion. Laboratory experiments with riparian substrates overlain with nitrate enriched solutions revealed a loss of only 5–8% of the nitrate during 48 h incubation at 12°C. However, 22–24% of the initial nitrate was depleted between 24 and 48 h when a second set of substrate cores was incubated at 20°C. Short-term (3 h) incubations of fresh substrates amended with acetylene were used to estimate in situ denitrification potentials which varied from 0.05–3.19 g N g^–1 d^–1 for organic and sandy sediments. Denitrification potentials were highly correlated with initial nitrate content of substrate samples implying that low nitrate levels in ground water and riparian substrates may be an important factor in controlling denitrification rates. The efficiency of nitrate removal in spring-fed drainage lines is also limited by short water residence times of < 1 h within the riparian zone. These data suggest that routes of ground water movement and substrate characteristics are important in determining nitrate depletion within stream riparian areas.     

A dye release assay for determination of lysostaphin activity

We describe a method for determination of lysostaphin activity using Remazol Brilliant Blue R (RBB)-dyed staphylococcal cells or RBB-dyed staphylococcal peptidoglycan as substrate. The dyed substrates are easy to prepare and are stable for at least 6 months. Soluble hydrolytic products released by lysostaphin are measured spectrophotometrically at 595 nm after the insoluble substrate is removed by filtration or centrifugation. The dye release assay is more sensitive and more accurate than the previously described turbidimetric assay.     

Deoxyribose ring conformation of [d(GGTATACC)]2: an analysis of vicinal proton-proton coupling constants from two-dimensional proton nuclear magnetic resonance

Exchangeable and nonexchangeable protons of [d(GGTATACC)]2 in aqueous cacodylate solution were assigned from two-dimensional nuclear Overhausser effect (2D NOE) spectra. With phase-sensitive COSY and double quantum filtered COSY (DQF-COSY) experiments, the cross-peaks resulting from deoxyribose ring conformation sensitive proton-proton vicinal couplings, i.e., all 1'-2', 1'-2", 2'-3', and 3'-4' couplings and six from 2"-3' couplings, were observed. From the cross-peak fine structure, the 2',2" proton assignments can be confirmed; coupling constants J1'2' and J1'2" and sums of coupling constants involving H2' and H2" for all residues and H3' for C8 were obtained. The DISCO procedure [Kessler, H., Muller, A., & Oschkinat, H. (1985) Magn. Reson. Chem. 23, 844-852] was used to extract individual 1'-2' and 1'-2" coupling constants. The sum of coupling constants involving H1' or H3' was measured from the one-dimensional spectrum where signal overlap is not a problem. Analysis of the resulting coupling constants and sums of coupling constants, in the manner of Rinkel and Altona [Rinkel, L. J., & Altona, C. (1987) J. Biomol. Struct. Dyn. 4, 621-649], led to the following conclusion: C2'-endo deoxyribose ring conformation is predominant for every residue, but a significant amount of C3'-endo conformation may exist, ranging from 14% to 30%.     

Comparison of activity and conformation changes during refolding of urea-denatured creatine kinase

The course of the recovery of the enzymatic activity and the native conformation during the renaturation of urea-denatured creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) has been studied. Under suitable conditions, an activity recovery of 95% can be obtained and the reactivation follows a triphasic course. The initial two phases are relatively fast, whereas the slow phase takes some 24 h to reach completion. The recovery of the native conformation has been followed by changes in fluorescence, ultraviolet absorption and in exposed SH groups and has been shown to be a biphasic process. Both the reactivation and the refolding processes are independent of protein concentrations within a certain range, showing that the dimerization of the enzyme molecule is not rate-limiting. A comparison of the rate constants for the refolding of the molecule with those for the recovery of its catalytic activity shows that these are not synchronized and the activity recovery approaches completion after the refolding and dimerization of the subunits so far as can be detected by the methods employed. The final stage of refolding with complete activity recovery probably involves subtle conformational changes of the dimeric enzyme molecule not detectable by the physiochemical methods used in the present study.     

钙通道阻断剂异搏定对缺氧性肺动脉高压形成的影响

为考察Ca~( )在缺氧性肺动脉高压形成中的作用,我们观察了钙通道阻断剂异搏定对慢性连续性缺氧大鼠肺动脉压及左右心功能的影响。将动物置于模拟海拔5000m高原的低压舱内,腹腔注射异搏定,剂量为4mg/kg BW,每日两次。实验结果表明:异搏定可以减弱缺氧15天所引起的肺动脉压升高和右心功能加强的程度,对颈动脉压及左心功能无明显影响,提示Ca~( )的跨膜内流是构成缺氧性肺动脉高压形成的重要基础之一。我们还比较了异搏定对缺氧持续时间不同(15天、10天、5天)的大鼠肺循环的影响,并讨论了异搏定发生作用的机制。     

血友病患者血清中淋巴腺病病毒/人T细胞Ⅲ型病毒抗体检测

对浙江省1982～1984年注射了美国产浓缩Ⅶ因子制剂的18例血友病患者,用酶联免疫吸附法(ELTSA)检测了血清中淋巴腺病病毒/人T细胞Ⅲ型病毒(LAV/HTLV-Ⅲ)抗体,发现4例阳性,并经免疫荧光试验和Western印迹法证实。2例应用了国产浓缩Ⅷ因子者抗体阴性。一例从美国来华旅游死于艾滋病者,LAV/HTLV-Ⅲ抗体阳性。本研究证明,LAV/HTLVⅢ病毒巳通过美国生产的Ⅷ因子制剂传入中国。     

外胚层细胞诱导后的间隙连接

用电镜观察了经受诱导作用之后胚胎细胞的冷冻蚀刻复型膜。和未经诱导作用的对照组比较,早期和中期神经胚的神经上皮细胞以及经过豚鼠骨髓粗提液(BME)——一种有效的异源中胚层诱导物——处理过的早期原肠胚外胚层,它们的间隙连接都处于活跃的动态状态。用图像分析仪测得的间隙连接连接子的排列密度,指出经受过诱导作用的三组分别和未经受诱导作用的对照组比较,计算求出P值,神经上皮两组和对照组的差别为非常显著,BME处理过的细胞和对照组的差别为显著。结合对照组与诱导后胚胎细胞间隙连接连接子的变化讨论了它们在信息传递上可能起的作用。     

An essential tryptophan residue for rabbit muscle creatine kinase

The tryptophan residues in rabbit muscle creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) have been modified by dimethyl(2-hydroxy-5-nitrobenzyl) sulfonium bromide after reversible protection of the reactive SH groups. The modification of two tryptophan residues as measured by spectrophotometric titration leads to complete loss of enzymatic activity. Control experiments show that reversible protection of the reactive SH groups as S-sulfonates followed by reduction results in nearly quantitative recovery of enzyme activity. The presence of a 410 nm absorption maximum and the decrease in fluorescence of the modified enzyme indicate the modification of tryptophan residues. At the same time, SH determinations after reduction of the modified enzyme show that the reagent has not affected the protected SH groups. Quantitative treatment of the data (Tsou, C.-L. (1962) Sci. Sin. 11, 1535 1558) shows that among the tryptophan residues modified, one is essential for its catalytic activity. The presence of substrates partially protects the modification of tryptophan residues as well as the inactivation, suggesting that the essential tryptophan residue is situated at the active site of this enzyme.     

Regional Distribution of Calmodulin Activity in Rat Brain

Calmodulin activity in 68 discrete areas of rat brain, obtained by micropunch technique, was assessed by its capacity to activate a calmodulin-sensitive form of phosphodiesterase. In general, the activity of calmodulin was higher in the telencephalon, limbic system, and hypothalamus than in the mesencephalon, pons, cerebellum, and medulla. However, there were substantial differences in calmodulin activity in discrete nuclei of each region. The regional distribution of calmodulin activity in rat brain does not appear to correlate with that of any of the known putative neurotransmitters or peptides.