Modulation of immobilized enzyme activity by altering the hydrophobicity of nylon-grafted membranes: Part 2: Non-isothermal conditions

Lactose hydrolysis by β-galactosidase immobilized on two nylon membranes, differently grafted, has been studied in a bioreactor operating under isothermal and non-isothermal conditions. One membrane (M₁) was obtained by chemical grafting of methylmethacrylate (MAA); the other one (M₂) by a double chemical grafting: styrene (Sty) and MAA. Hexamethylenediamine was used as a spacer between the grafted membranes and the enzyme. Both membranes have been physically characterized studying their permeabilities in presence of pressure or temperature gradients. Under non-isothermal conditions, the increase in activity of membrane M₂ was higher than that of membrane M₁. The and β coefficients, giving the percentage of activity increase when a temperature difference of 1°C is applied across the catalytic membranes, have been calculated. Results have been discussed with reference to the greater hydrophobicity of membrane M₂ with respect to membrane M₁, the hydrophobicity being a prerequisite for the occurrence of the process of thermodialysis.     

有机肥氮投入比例对双季稻田根际土壤微生物生物量碳、氮和微生物熵的影响

为探明不同有机肥氮素占总氮投入的百分比对双季稻区早、晚稻各生育时期稻田根际土壤微生物的影响,本研究以大田定位试验为平台,应用氯仿熏蒸-K₂SO₄提取法和化学分析法系统分析了施用化肥N(M₁)、30%有机肥N(M₂)、50%有机肥N(M₃)、100%有机肥N(M₄)和无N对照(M₀)5个不同施肥处理双季稻田根际土壤微生物生物量碳(MBC)、微生物生物量氮(MBN)和微生物熵的差异.结果表明: 在早稻和晚稻各主要生育时期,施肥措施均能提高稻田根际土壤MBC、MBN和微生物熵,各施肥处理根际土壤MBC、MBN和微生物熵均随水稻生育期推进呈先增加后降低的变化趋势,均于齐穗期达到最大值,成熟期为最低值;其中,各处理双季稻田根际土壤MBC、MBN、MBC/MBN值和微生物熵一般均表现为M₄＞M₃＞M₂＞M₁＞M₀,M₂、M₃和M₄处理间均无显著差异,但均显著高于M₀处理.可见,单独施用化肥措施对提高根际土壤微生物生物量碳、氮和微生物熵效果有限,施用有机肥或有机无机肥配施提高根际土壤微生物生物量碳、氮和微生物熵的效果较好.     

Effects of JO 1784, A selective sigma ligand, on the autoradiographic localization of M2 and M2 muscarinic receptor subtypes in trimethyltin treated rats

The distribution patterns of M₁ and M₂ muscarinic receptor subtypes following TMT and JO 1784 administration in the male Sprague-Dawley rat were investigated. In the present study, JO 1784 was injected in doses of 1, 4 and 16 mg/kg i.p. for one week prior to the single injection of TMT (8 mg/kg i.p.) and subsequently for 33 days. The effects of JO 1784 on the density of muscarinic receptor sub-types (M₁ and M₂) in the control and trimethyltin (TMT) treated rats were then evaluated. The topographic distribution and changes in muscarinic (M₁ and M₂) receptor densities were determined by means of autoradiography using [³H]quinuclidinylbenzilate (QNB). Both sub-types of muscarinic receptors contributed to the observed decrease in total muscarinic receptor binding in TMT-treated rats. In control rats, JO 1784 alone decreased M₁ receptor density in the amygdaloid nuclei, basal ganglia, cortex and hippocampus and decreased M₂ receptor density in the amygdaloid nuclei, basal ganglia, cortex, hippocampus, hypothalamus and septal regions. In TMT treated rats, chronic JO 1784 administration has a “neuroprotective effect” on both M₁ and M₂ receptors subtypes. Thus, following chronic administration of JO 1784 to TMT treated rats, both increases and decreases in M₁ receptor density were observed relative to TMT animals. A significant increase in M₁ receptor density was found in the cortex, olfactory regions, septum, thalamus and basal forebrain nuclei. In the hippocampus (CA₂ and CA₃), a significant decrease in M₁ receptor density was observed. In TMT-treated rats, JO 1784 produced a significant increase in M₂ receptor density in several brain regions with the most marked effects occurring in the amygdaloid nuclei, basal ganglia, cortex, hippocampus and hypothalamus. The ability of the selective sigma ligand, JO 1784, to attenuate the loss of muscarinic receptors in TMT treated rats could be of importance in the development of novel neuroprotective drugs.     

Novel oxotremorine-related heterocyclic derivatives: Synthesis and in vitro pharmacology at the muscarinic receptor subtypes

A set of novel heterocyclic ligands (6–27) structurally related to Oxotremorine 2 was designed, synthesized and tested at muscarinic receptor subtypes (mAChRs). In the binding experiments at cloned human receptors (hm1–5), compounds 7 and 15 evidenced a remarkable affinity and selectivity for the hm2 subtype. The in vitro functional assays, performed on a selected group of derivatives at M₁, M₂, and M₃ tissue preparations, singled out the 3-butynyloxy-5-methylisoxazole trimethylammonium salt 7 as a potent unselective muscarinic agonist [pEC₅₀: 7.40 (M₁), 8.18 (M₂), and 8.14 (M₃)], whereas its 5-phenyl analogue 12 behaved as a muscarinic antagonist, slightly selective for the M₁ subtype [pK_B: 6.88 (M₁), 5.95 (M₂), 5.53 (M₃)]. Moreover, the functional data put in evidence that the presence of the piperidine ring may generate a functional selectivity, e.g., an M₁ antagonist/M₂ partial agonist/M₃ full agonist profile (compound 21), at variance with the corresponding quaternary ammonium salt (compound 22) which behaved as a muscarinic agonist at all M_1–3 receptors, with an appreciable selectivity for the cardiac M₂ receptors.     

长期不同施肥对玉米根茬生物量及养分累积量的影响

以黄土高原南部两个长期定位试验（分别开始于1990和2003年）为研究对象,探讨了不同肥料处理对玉米根茬生物产量和养分累积的影响.于2011年10月玉米收获后采集0～20 cm土层不同施肥处理玉米根茬.结果表明：与不施肥及偏施N、NK、PK化肥相比,氮磷配施（NP）、氮磷钾平衡施肥（NPK）、有机无机配施（M₁NPK、M₂NPK）及化肥配合秸秆（SNPK）处理均显著提高了玉米根茬干质量.根茬固碳量及氮、磷、钾养分累积量在NP、NPK、M₁NPK、M₂NPK、SNPK处理显著高于不施肥和偏施N、NK、PK化肥处理,其中以有机无机配施处理效果最好.与不施氮肥（N₀）相比,施氮120 kg N·hm^-2（N₁₂₀）和240 kg N·hm^-2（N₂₄₀）处理根茬干质量分别提高38%和45%,高量氮肥对根茬增量效果不显著.施用氮肥也显著提高了根茬碳、氮、磷、钾累积量.根茬可溶性有机碳、可溶性总氮含量在NP、NPK、M₁NPK、M₂NPK、SNPK及N₁₂₀和N240处理中较高.氮磷钾平衡施肥、有机无机配施以及秸秆还田处理降低了根茬的纤维素、木质素含量.根茬C/N、木质素/N在CK、PK、N₀处理间显著高于其他施肥处理.因此,氮磷配施、氮磷钾平衡施肥、有机无机配施及秸秆还田处理能够促进玉米根生长,提高营养成分含量,有利于土壤培肥和固碳.     

The neuroprotective effect of tacrine on trimethyltin induced memory and muscarinic receptor dysfunction in the rat

In this study chronic (39 days) tacrine (3 mg/kg i.p.) treatment significantly improved trimethyltin (8 mg/kg i.p.) induced deficits in spatial navigation. Tacrine also reduced trimethyltin induced hyperactivity and passive avoidance deficits but these effects did not reach statistical significance. The effect of trimethyltin on muscarinic (M₁ and M₂) receptor sites was determined by means of quantitative autoradiography using [³H]quinuclidinyl benzilate. A selective pattern of M₁ and M₂ receptor loss was observed mainly affecting the hippocampus and other limbic structures while leaving other brain regions intact. Tacrine successfully prevented the M₁ and M₂ receptor loss in the CA₁ and CA₁ hippocampal subfields. The improvement in trimethyltin behavioural toxicity following tacrine treatment may be related to the protective effect of this compound on muscarinic receptor density in the hippocampal formation and lends support to the hypothesis that cholinergic system dysfunction may be primarily responsible for trimethyltin induced deficits in cognitive function.     

Synthesis, NMR spectroscopy study, and antimuscarinic activity of a series of 2-(Acyloxymethyl)-1,3-dioxolanes

A series of 1,3-dioxolane-based ligands, bearing hydroxymethyl or ester functionalities, was synthesized and tested as potential muscarinic antagonists. The compounds display moderate to low affinity for the three receptor subtypes M₁-M₃, with some of them showing a significant selectivity for the M₃ subtype. The configurational and conformational properties were studied using NOE experiments and vicinal coupling constants. The ¹H and ¹³C NMR chemical shifts show stereochemically dependent trends. Quantitative analysis of conformer populations showed that the exocyclic CH₂N⁺(CH₃)₃ group is prevalently in a pseudo-axial orientation in the cis isomers and in a pseudo-equatorial orientation in the trans isomers.     

有机物沟埋还田模式与花后灌水量对玉米光合特性和产量的影响

有机物沟埋还田与花后灌水配合对增加玉米田保水供水能力,提高玉米花后光合性能、实现节水增产有重要意义.本试验以郑单958为供试材料,设置有机物沟埋还田和花后灌水量两个因素,有机物沟埋还田包括不还田(M₀)、秸秆单还田(M₁)和牛粪秸秆混合还田(M₂)3个还田类型,花后灌水量包括450 mm(W₁)和325 mm(W₂)2个水平,研究了其对玉米穗位叶光合性能、光系统Ⅱ(PSⅡ)效率和产量等的影响.结果表明:与秸秆单还田比较,牛粪秸秆混合还田有效提高了玉米花后光合能力和各器官的干物质积累量;与节水灌溉相比,正常灌溉加强了有机物还田对玉米光合能力的促进作用.牛粪秸秆混合还田与正常灌溉结合可显著提高玉米花后叶片的光合速率(P_n)、气孔导度(g_s)和蒸腾速率(T_r),降低胞间CO₂浓度(C_i);提高玉米花后叶片PSⅡ的最大光化学效率(φ_po)和捕获的激子将电子传递到电子传递链中Q_A下游的电子受体的概率(Ψ_o);改善花后叶片光能利用率,维持花后叶片较高的光合性能;同时增加花后玉米各器官干物质的量,提高干物质总积累量和转运能力,有利于花后同化物对籽粒的分配,最终获得高产.节水灌溉降低了叶片的光合性能,造成产量的下降;但配合牛粪秸秆混合还田与不还田处理相比,水分利用效率、籽粒增长速率和增产效果均优于正常灌水.这表明牛粪秸秆混合还田与正常灌溉结合可有效提高玉米花后光合性能,增加干物质积累量,促进玉米增产;牛粪秸秆混合还田与节水灌溉结合一定程度上降低了因减少灌溉量造成的减产.     

Resonance raman spectroscopy of chemically modified and isotopically labelled purple membranes. II. Kinetic studies

Kinetic resonance Raman spectra of native and isotopically labelled purple membranes are compared. Using these data and the assignments of the previous paper in this sequence, we have confirmed that the Schiff base is deprotonated at times that are short in comparison to M₄₁₂ evolution. In addition, by monitoring the kinetic resonance Raman spectra in ²H₂O with 488.0 nm excitation we have been able to characterize in more detail the vibrational features associated with this unprotonated intermediate that precedes M₄₁₂. Furthermore, the kinetic spectra of fully deuterated purple membranes in H₂O have allowed us to assign the 1465 cm⁻¹ band in these spectra to the C=C stretching frequency of BR₅₇₀ and the 1512 cm⁻¹ band to the C=C stretching frequency of M₄₁₂. These spectra have also provided an indication of a Raman spectral feature associated with O₆₄₀ and, finally, our kinetic spectra have provided evidence that there is a significant alteration in the rate constants for the evolution of the various intermediates when the non-exchangeable protons on the membrane are replaced by deuterons.     

小家鼠(Mus musculus)种群动态预测及机制的探讨

我国学者夏武平(1966)用9月铗日捕获率预测东北棕背(鼠平)(Clethrionomys rufocanus)数量以来,严志堂等(1983)应用种群年龄结构的回归方程预测新疆天山北麓农区塔西河地区的小家鼠(Mus musculus)种群密度获初步成功,并根据该文提供的一个改进公式M₁₀=5.60x+5.80(x表示种群年龄的成亚比,M₁₀表示小家鼠10月数量,即年内最高数量),提前11个月预测该地的小家鼠种群最高数量。     

植物生长调节剂对银柴胡细胞生长特性的影响

利用细胞悬浮培养技术,研究不同激素对银柴胡细胞生长特性及过氧化氢酶和硝酸还原酶活性的影响。结果表明,第4种激素配比的培养液M₄(MS+6-BA 1.0 mg·L^-1+2,4-D 1.0 mg·L^-1+NAA 1.0 mg·L^-1)的细胞鲜重、干重,细胞数目均明显高于其他处理;M₄处理可有效增强银柴胡细胞硝酸还原酶的活性,提高氮素利用率;M₂(MS+6-BA 1.0mg·L^-1+NAA0.5 mg·L^-1)处理有利于增强银柴胡细胞过氧化氢酶活性。     

MRI study of the inhibitory effect of new spin traps on in vivo CCl4-induced hepatotoxicity in rats

Acute CCl₄ hepatotoxicity is thought to occur as a result of free generated from the metabolism of CCl₄ in the liver. With the use of MRI it is possible to detect in vivo a CCl₄-induced edematous region surrounding the major branch of the hepatic portal vein in the right lobe. Inhibition of the CCl₄-induced response has been obtained by pretreatment with the spin trap, PBN, 30 min prior to CCl₄ exposure. The inhibitory effect of two new traps, M₃PO or methyl-DMPO, and PhM₂PO or phenyl-DMPO, on in vivo CCl₄-induced acute hepatotoxicity was investigated. Both PhM₂PO and M₃PO were found to inhibit the CCl₄-induced response at lower concentrations (0.35 M/kg body weight) than PBN (0.70 M/kg body weight). However, both M₃PO and PhM₂PO were also found to induce and edematous response at the same concentrations used for the PBN studies (0.70 M/kg body weight). PhM₂PO, at a concentration of 0.35 M/kg body weight, was 93% as efficient as PBN, at a concentration of 0.70 M/kg body weight; whereas M₃PO, at a concentration of 0.35 M/kg, was 89% as efficient as PBN at 0.70 M/kg body weight. Electron micrographs were obtained from small liver sections taken in proximity to the major branch of the hepatic portal veins of all treatment groups. The electron microscopy investigations support the MRI findings.     

6-(4-Phenyl-benzyloxy-methyl) guvacine. Synthesis, GABA uptake inhibitor and muscarinic properties

6-(4-Phenyl-benzyloxy-methyl) guvacine was synthesized. Surprisingly the compound was devoid of the γ-aminobutyric acid (GABA) uptake inhibitory activity of its parent compound guvacine, but instead showed affinities for the muscarinic M₁ and M₂ receptors.     

The effects of carbon dioxide concentration on oxygen evolution and fluorescence transients in synchronous cultures of Chlorella pyrenoidosa

1. 1. The steady-state fluorescence yield of Chlorella pyrenoidosa is strongly affected by CO₂ concentration: the yield is approximately 2-fold higher in the presence than in the absence of CO₂. During induction, in the presence of saturating CO₂, accelerating oxygen evolution is paralleled by rising fluorescence (M₂-P₃ transient); in the absence of CO₂, fluorescence yield remains at the low M₂ level.

2. 2. Both illumination and CO₂ content are important in determining the steady-state fluorescence yield: at lower illuminations, lower concentrations of CO₂ are required to obtain a maximum fluorescence yield.

3. 3. The slow fluorescence transients are not affected directly by pH but only indirectly through the CO₂ concentration.

4. 4. The CO₂-dependent fluorescence rise (M₂-P₃ transient) is most readily observed in cells harvested early in the light period of a synchronous culture, but it can also be elicited in cells harvested during the dark period.

5. 5. Addition of 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU) to CO₂-deprived cells raises the fluorescence yield approximately 4-fold, that is to the same high level as cells supplied with CO₂ and DCMU.

6. 6. The effects of CO₂ provide a new example of a marked parallelism between photosynthetic electron transport and fluorescence. To explain such parallelism, it seems necessary to postulate large changes in the de-excitation processes within Photosystem II units or in the distribution of excitation between Photosystems I and II.

Metal–salt co-tolerance and metal removal by indigenous cyanobacterial strains

Chromium and salt tolerance in five indigenous cyanobacterial strains isolated from contaminated sites was investigated along with their metal bioaccumulative potential. All the five species showed significantly better growth when the medium was spiked with salt or chromium. As compared to single metal or salt treatment, the binary metal–salt (MS) treatments had more favorable effect on cyanobacterial growth as indicated by significantly higher concentration of the primary photosynthetic pigment chlorophyll at M₂₀S₂₀₀₀ (9.9–25.3 μg/mL) as compared to that at M₀S₀ (4.0–12.3 μg/mL). Similarly biomass was much higher at M₂₀S₁₀₀₀ and M₂₀S₂₀₀₀ (41.8–86.2 mg/10 mL) as compared to that at control, M₀S₀ (21.5–36.3 mg/10 mL). Accessory pigments like carotenoids and phycobilinproteins too tended to increase significantly in response to both metal and salts in the two species of Lyngbya (L. putealis and L. ceylanica var. constricta) and Gloeocapsa. These species also showed greater potential of chromium bioaccumulation, which increased further as both salt and metal concentration increased. In the two species of Nostoc however, bioaccumulative potential improve at higher metal concentration, but not affected significantly by salt concentration.     

盐胁迫对VA菌根形成及接种VAM真菌对植物耐盐性的效应

用盆栽法研究了不同土壤含盐量条件下接种不同VAM真菌菌株对棉花、玉米、大豆和甜瓜耐盐性的影响。结果表明,随着土壤中NaCl水平提高,真菌对棉花、玉米、大豆和甜瓜的侵染率下降,其生长量亦均呈递减趋势但对VAM真菌的依赖性则呈明显递增趋势。接种VAM真菌促进了棉花、玉米和大豆的生长,如在NaCl施入量为0～3g·kg^-1时,接种M₁菌株使棉花干重提高4.6～80.9%;在NaCl施入量为0～2.5g·kg^-1时,接种M₁菌株的玉米干重比对照增加20～109.6%;NaCl施入量为1g·kg^-1时,接种M₁和M₂两菌株使大豆干重分别增加22.1%和10.2%.不同的VAM真菌菌株对同一植物的耐盐性以及同一种VAM真菌菌株对不同植物的耐盐性的影响程度不同。可以认为,VAM真菌与植物共生对植物在盐渍环境中的生存起着重要的作用;接种VAM真菌可以提高植物在盐渍土壤上的生产能力并减轻植物因盐害造成的产量损失。     

长期施肥对土壤氨氧化微生物的影响

长期施肥可改变土壤碳氮等养分供应,进而影响微生物数量与群落组成。本研究基于棕壤长期定位实验站,分析不同施肥方式下(不施肥,CK;低量无机氮肥,N₂;高量无机氮肥,N₄;有机无机氮肥配施,M₂N₂)土壤氨氧化古菌(AOA)和细菌(AOB)的变化,为土壤氮素转化的微生物学机制和培肥土壤提供依据。结果表明:不同施肥方式下,土壤AOA与AOB的数量比值为2.28～61.95。与CK相比,施肥后土壤AOA数量降低了1.6%～13.6%。N₄处理AOB数量随土壤深度增加呈先降低后升高的趋势,其他处理则相反。土壤AOB群落Shannon多样性指数、均匀度指数和Simpson指数均高于AOA。M₂N₂处理0～20 cm土层土壤AOB多样性增加,但AOA多样性降低。土壤AOB主要因土壤深度发生聚类,AOB和AOA均未因施肥方式发生聚类。综上,长期施肥改变了土壤AOA和AOB的构成状况,AOA对环境变化较为敏感,AOB较为丰富和稳定。     

水稻两性生殖细胞的N-甲基-N-亚硝基脲诱变方法

N-甲基-N-亚硝基脲(MNU)被用于水稻(Oryza sativa)受精卵的诱变。通过水稻辽盐6号成熟生殖器官的MNU体内同步处理及后代群体筛查, 确立了水稻两性生殖细胞的MNU诱变方法。与辽盐6号受精卵的MNU处理相比, 各组条件下两性生殖细胞的MNU处理明显使M₁群体生长发育的指标降低及M₁-M₂群体中突变性状的发生率升高。两性生殖细胞在含有1.5 mmol?L ^-1 MNU和10 mmol?L ^-1 PO₄ ^3-的缓冲液(pH4.8)中处理60分钟, 突变性状发生率是基于受精卵MNU处理的3倍。进一步筛查M₃群体, 获得了包含新型植株和籽粒突变体的纯合突变体系列。研究结果表明, 水稻两性生殖细胞的MNU诱变可显著提高广谱诱变效率。该技术的应用可为水稻的未知功能基因鉴定和育种所需的各种突变体规模化开发提供高效的技术支撑。     

水稻两性生殖细胞的N-甲基-N-亚硝基脲诱变方法

N-甲基-N-亚硝基脲(MNU)被用于水稻(Oryza sativa)受精卵的诱变。通过水稻辽盐6号成熟生殖器官的MNU体内同步处理及后代群体筛查, 确立了水稻两性生殖细胞的MNU诱变方法。与辽盐6号受精卵的MNU处理相比, 各组条件下两性生殖细胞的MNU处理明显使M₁群体生长发育的指标降低及M₁-M₂群体中突变性状的发生率升高。两性生殖细胞在含有1.5 mmol∙L ^-1 MNU和10 mmol∙L ^-1 PO₄ ^3-的缓冲液(pH4.8)中处理60分钟, 突变性状发生率是基于受精卵MNU处理的3倍。进一步筛查M₃群体, 获得了包含新型植株和籽粒突变体的纯合突变体系列。研究结果表明, 水稻两性生殖细胞的MNU诱变可显著提高广谱诱变效率。该技术的应用可为水稻的未知功能基因鉴定和育种所需的各种突变体规模化开发提供高效的技术支撑。     

Adult human brain expresses four different molecular forms of neural cell adhesion molecules

Using antibodies to rat neural cell adhesion molecules (NCAM), we analyzed the NCAM of adult human brain. Various regions of the brain were analyzed quantitatively by Western blot. Grey matter showed four bands of NCAM with apparent molecular weights of 180,000, 170,000, 140,000 and 120,000. White matter showed one major band with an apparent M_r of 120,000 and a minor band of 180,000. Cerebellar grey matter contained mainly 170,000, 140,000 and 120,000, white cerebellar white matter had only 180,000 and 120,000 M₁ NCAMS. Spinal cord showed mainly 120,000 M_r NCAM. Deglycosylation using N-glycanase resulted in 170,000, 160,000, 130,000 and 110,000 M_r proteins, suggesting that the four forms of human NCAM are derived from individual polypeptides. The presence of 170,000 M₁ NCAM is unique to human brain.