Effect of Inorganic and Organic Tin Compounds on ACh- and Voltage-Activated Na Currents

1. Inorganic tin and organotin compounds, occurring in aquatic ecosystems, are toxic and can cause behavioral abnormalities in living organisms. To determine the possible neuronal basis of these actions, the effects of both forms of Sn were studied on identified neurones of the mollusk, Lymnaea stagnalis L.2. SnCl₂ caused a dose-dependent decrease in the acetylcholine (Ach)-induced inward current. The effective threshold concentration, measured by a two microelectrode voltage clamp technique, was 0.1 M, and the maximal effect occurred at 5 M SnCl₂. The depression of the inward current was greater after a 10 min preapplication (20%) than after 3 min treatment (7%).3. The next series of experiments compared the actions of inorganic or organic tin compounds. In whole cell clamp experiments both (CH₃)₂SnCl₂ and (CH₃)₃SnCl, like inorganic Sn, decreased the amplitude of Ach-induced current. Increasing the duration of the preapplication time resulted in an increase in the effect, but the action was not reversible. SnCl₂ treatment caused a concentration-dependent alteration (initial potentiation followed by depression) of the amplitude of I _Na(V) over the whole voltage range and slightly shifted the I–V curves to the left. In contrast, trimethyl tin decreased the amplitude of I _Na(V) only at high concentration (100 M). The activation time course of I _Na was increased ( = 0.43 ms in control and 0.55 ms in Sn), but Sn did not alter the inactivation parameters ( = 3.43 and 3.41 ms).4. These results support earlier findings that agonist- and voltage-activated channels are direct targets of toxic metals. We conclude that tin in both inorganic and organic forms acts at neuronal membranes to modulate synaptic transmission through direct actions on agonist-activated ion channels, and suggest that these actions may be the basis of the altered behavior of animals in tin-polluted environments.     

Multicolour Fluorescence Imaging of Sugar Beet Leaves with Different Nitrogen Status by Flash Lamp UV-Excitation

Fluorescence images of leaves of sugar beet plants (Beta vulgaris L. cv. Patricia) grown on an experimental field with different fertilisation doses of nitrogen [0, 3, 6, 9, 12, 15 g(N) m^–2] were taken, applying a new multicolour flash-lamp fluorescence imaging system (FL-FIS). Fluorescence was excited by the UV-range (280–400 nm, _max = 340 nm) of a pulsed Xenon lamp. The images were acquired successively in the four fluorescence bands of leaves near 440, 520, 690, and 740 nm (F₄₄₀, F₅₂₀, F₆₉₀, F₇₄₀) by means of a CCD-camera. Parallel measurements were performed to characterise the physiological state of the leaves (nitrogen content, invert-sugars, chlorophylls and carotenoids as well as chlorophyll fluorescence induction kinetics and beet yield). The fluorescence images indicated a differential local patchiness across the leaf blade for the four fluorescence bands. The blue (F₄₄₀) and green fluorescence (F₅₂₀) were high in the leaf veins, whereas the red (F₆₉₀) and far-red (F₇₄₀) chlorophyll (Chl) fluorescences were more pronounced in the intercostal leaf areas. Sugar beet plants with high N supply could be distinguished from beet plants with low N supply by lower values of F₄₄₀/F₆₉₀ and F₄₄₀/F₇₄₀. Both the blue-green fluorescence and the Chl fluorescence rose at a higher N application. This increase was more pronounced for the Chl fluorescence than for the blue-green one. The results demonstrate that fluorescence ratio imaging of leaves can be applied for a non-destructive monitoring of differences in nitrogen supply. The FL-FIS is a valuable diagnostic tool for screening site-specific differences in N-availability which is required for precision farming.     

The L6 membrane proteins--a new four-transmembrane superfamily

L6, IL-TMP, and TM4SF5 are cell surface proteins predicted to have four transmembrane domains. Previous sequence analysis led to their assignment as members of the tetraspanin superfamily. In this paper, we identify a new sequence (L6D) that is strikingly similar to L6, IL-TMP, and TM4SF5. Analyses of these four sequences indicate that they are not significantly related to genuine tetraspanins, but instead constitute their own L6 superfamily.     

冬小麦原生质体培养的胚状体直接发生

冬小麦品种“京花一号”胚性愈伤组织在改良的N₆培养基(NBD培养基)上继代得到易碎型胚性愈伤组织,转入改良MS液体培养基(MSDL培养基)后得到胚性悬浮系,分离的原生质体在改良的MS培养基(MSDP培养基)上培养,再生细胞直接产生体细胞胚胎,并再生出完整植株。体细胞胚胎形成过程与小麦合子胚的形成过程十分相似。     

灭活的双歧杆菌对肠上皮细胞粘附及其影响因素的研究

目的 观察灭活的青春双歧杆菌对人大肠癌细胞系ＣＣＬ－２２９的粘附以及影响粘附的因素。方法 通过与双歧杆菌活菌比较,灭活的双歧杆菌同样能粘附于肠上皮细胞,并且耗尽培养上清有利于双歧杆菌粘附。结果 粘附具有显著的浓度效应;粘附效果与孵育环境的ｐＨ值有关;高温处理耗尽培养上清对粘附无明显影响。结论 灭活的双歧杆菌可能具有与活菌相同的生态效应。     

银杏种子和叶的蛋白质分析

采用分级提最方法提取银杏（Ginkgo biloba L.）种子和银杏叶蛋白质,并进行各组分蛋白质含量测定和银杏种子后熟过程中蛋白质含量动态变化的分析。结果表明,银杏种子以水溶性和盐溶性蛋白质为主,银杏叶以醇溶性蛋白质为主。银杏叶中主要蛋白质在HPLC柱中的保留时间为3.457min,相对含量达70%以上。银杏叶蛋白质含有丰富的亮氨酸、缬氨酸、赖氨酸和色氨酸,其含量分别为20.23%、13.35%、4.81%和3.73%。萌发种子胚体中的蛋白质主要是醇溶性蛋白质和谷蛋白类蛋白质。在种子萌发过程中,胚乳蛋白质含量明显增加,播种后第3周和萌发时总蛋白质含量达到高峰。     

Retinoic Acid and Its Geometrical Isomers Block Both Growth and Fusion of L6 Myoblasts by Modulating the Expression of Protein Kinase A

All-trans retinoic acid (RA) and its geometrical isomers, such as 9-cis RA, 13-cis RA, and 9,13-di-cis RA, strongly inhibited both growth and fusion of L6 myoblasts. However, illumination of white light diminished their inhibitory activity on membrane fusion with little effect on cell growth. During myogenic differentiation, the intracellular level of cAMP decreased whereas the total activity of protein kinase A as well as the protein level of its regulatory subunit I (RI) and catalytic subunit (C) increased. RAs raised the intracellular level of cAMP by over 3-fold, but decreased the total activity of protein kinase A. Like RAs, dibutyryl-cAMP inhibited myoblast fusion and reduced the expression of both RI and C subunits. These results suggest that RAs negatively modulate the differentiation of L6 myoblasts by increasing the intracellular level of cAMP, which may in turn down-regulate the expression of protein kinase A and hence its activity.     

Extractive cultivation of Lactococcus lactis using a polyethylene glycol/MgSO4 · 7H2O aqueous two-phase system to produce nisin

To relieve lactic acid inhibition, an aqueous two-phase system (ATPS) was used to grow Lactococcus lactis. Its composition was 11% (w/v) PEG 20000/3.5% (w/v) MgSO₄ 7H₂O. In this ATPS medium, the cells were completely partitioned in the bottom phase, and lactic acid had the biggest partition coefficient of the eight ATPS media tested. The cell biomass in this medium was 0.64 mg ml^–1, only 60% of that of the control medium, but nisin production (803 IU ml^–1) was enhanced by 33%. The increase in nisin was explained as a result of extraction of lactic acid from the bottom phase to the top one. The changes of tie-line length and phase volume ratio for the identical tie line could affect cell growth and nisin accumulation.     

Photosynthetic acclimation of higher plants to growth in fluctuating light environments

This paper describes a study into the potential of plants to acclimate to light environments that fluctuate over time periods between 15 min and 3 h. Plants of Arabidopsis thaliana (L.) Heynh., Digitalis purpurea L. and Silene dioica (L.) Clairv. were grown at an irradiance 100 mol m^-2 s^-1. After 4–6 weeks, they were transferred to light regimes that fluctuated between 100 and either 475 or 810 mol m^-2 s^-1, in a regular cycle, for 7 days. Plants were shown, in most cases, to be able to undergo photosynthetic acclimation under such conditions, increasing maximum photosynthetic rate. The extent of acclimation varied between species. A more detailed study with S. dioica showed that this acclimation involved changes in both Rubisco protein and cytochrome f content, with only marginal changes in pigment content and composition. Acclimation to fluctuating light, at the protein level, did not fully reflect the acclimation to continuous high light - Rubisco protein increased more than would be expected from the mean irradiance, but less than expected from the high irradiance; cytochrome f increased when neither the mean nor the high irradiance would be expected to induce an increase.This revised version was published online in October 2005 with corrections to the Cover Date.