The Casparian strip in pea epicotyls: Effects of light on its development

The Casparian strip, which is specific to roots, was studied in the epicotyls of dark-grown seedlings of pea (Pisum sativum L.) where it was found to have the same morphology and properties as the strip in roots. In dark-grown seedlings, the distance between the upper-most position of the Casparian strip and the bending point of the hook (about 37 mm) did not change during growth of the seedlings. In the uppermost 0.5-mm region of the region in which the Casparian strip could be detected by fluorescence microscopy, the plasma membrane was not firmly attached to the cell wall. The development of the Casparian strip continued for about 42 h after dark-grown seedlings were transferred to the light, indicating that (i) the cells that have been determined to form the Casparian strip in darkness form the strip in the light, and that (ii) it takes about 42 h for the cells to complete formation of the strip. Cells in the hook of dark-grown seedlings did not form a Casparian strip when such seedlings were transferred to the light. The Casparian strip was formed in rapidly elongating internodes of light-grown seedlings when the seedlings were transferred to darkness. Light did not control the formation of the Casparian strip in roots.Abbreviation PBS phosphate-buffered saline     

Effect of bicarbonate and root zone temperature on uptake of Zn,Fe, Mn and Cu by different rice cultivars (Oryza sativa L.) grown in calcareous soil

Pot experiments were conducted with a calcareous soil (Inceptisol) to elucidate the effects of bicarbonate (0 and 20 mM) and root zone temperature (15° and 25°C) on the uptake of Zn, Fe, Mn and Cu by "Zn-efficient" and "Zn-inefficient" rice cultivars. Bicarbonate decreased concentrations and total uptake of Zn in shoots of "Zn-inefficient" cultivars, especially of IR 26 at 25°C, but not in Zn-efficient cultivars. Bicarbonate decreased concentrations and uptake of Fe in shoots of Zn inefficient cultivars, particularly in IR 26. Concentrations and total uptake of Mn were lower in bicarbonate treatment in the Zn-inefficient cultivars at 15°C, and in all cultivars at 25°C. However, concentration and uptake of Cu were not affected by bicarbonate in all cultivars. Compared to the 25°C root zone temperature, the concentrations and total uptake of both Zn and Cu in shoots at 15°C were lower in Zn-inefficient than in the Zn-efficient cultivars. The results indicate that Zn-efficiency in rice is causally related to high tolerance of plant to elavated bicarbonate concentrations in soil solution.     

Effect of selenium and protein deficiency on selenium and glutathione peroxidase in rats

Twenty-four weanling male Wistar rats were divided into four groups fed diets containing adequate or deficient levels of selenium (0.5 ppm [+ Se] or <0.02 ppm [−Se] and protein (15% [+Pro] or 5% [−Pro]), but adequate levels of all other nutrients for 4 wk to determine the effects of Se deficiency and protein deficiency on tissue Se and glutathione peroxidase (GSHPx) activity in rats. Plasma, heart, liver, and kidney Se and GSHPx were significantly lower in Se-deficient groups in relation to Se-sufficient groups. In Se-deficient groups, Se and GSHPx were significantly higher in −Se−Pro rats in heart, liver, and kidney. Data analysis showed that there were significant interaction effects between dietary Se and protein on Se and GSHPx of rats. It is assumed that under the condition of Se deficiency. a low level of protein may decrease Se and GSHPx utilization, increase GSHPx synthesis, and result in Se redistribution. This could account for high levels of Se and GSHPx in the −Se−Pro rats compared to −Se+Pro rats.     

人血红细胞酪氨酸蛋白磷酸酶（PTPP）的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质

人血红细胞胞浆部分经（ＮＨ_４）_２ＳＯ_４沉淀,ＤＥＡＥ－纤维素（ＤＥ５２）柱层析,磷酸纤维素柱层析（Ｐ１１）得到部分纯化的ＰＴＰＰ,产率：５．７％,提纯１０７５倍。以（３２）￣Ｐ－Ｔｙｒ－Ｐｏｌｙ（Ｇ_４：Ｔ）作底物,测得其表征Ｋｍ约为０．５－０．８μｍｏｌ／Ｌ,该酶的最适ｐＨ和最适温度分别为７．０－７．８及３７－４０℃。Ｚｎ￣（２＋）等二价金属离子及Ｎａ_３ＶＯ_４等酸根基团对其活性有明显的抑制作用;ＥＤＴＡ、甘油及ＤＴＴ、巯基乙醇等则对其有强烈激活作用;而氟化物、酒石酸等对ＰＴＰＰ活性基本无影响。此外,某些蛋白质、氨基酸、核苷酸及抗肿瘤药物等对ＰＴＰＰ活性也都有不同程度的影响。特别是一些ＰＫｓ及ＰＰｓ在体外对ＰＴＰＰ活性也具有不同的作用。     

Seed development in Pisum

The Ih and lh _i alleles have been shown previously to reduce the level of endogenous gibberellin A₁ (GA₁) in shoots of pea (Pisum sativum L.), resulting in a dwarf phenotype compared with the wild type, cv. Torsdag (Lh). In addition, plants homozygous for the lh ⁱ allele have reduced seed yield compared with Lh (tall, wild type) and lh (dwarf) plants. In this paper we show that the lh ⁱ mutation is expressed in developing seeds and pods. Comparison of GA levels in young shoots and developing seeds of genotypes lh and lh ⁱ demonstrates that the relative severity of the two mutations varies in different tissues. Homozygous h ⁱ seeds have reduced GA levels, weigh less, and are less likely to develop to maturity when compared with Lh seeds. However, fertilization of lh ⁱ plants with Lh pollen increases seed GA levels, seed weight and seed survival, indicating that an increase in seed GA levels due to the presence of the Lh allele can restore normal seed growth. Pods developing on self-pollinated lh ⁱ plants are shorter than pods on Lh (wild type) plants, although this may be an indirect effect of the increased seed abortion of lh _i plants. Based on these results we suggest that endogenous GAs play an important role in the development of seeds of P. sativum L.Abbreviations GA_(n) gibberellin A_n We wish to thank Katherine McPherson, Peter Newman, Leigh Johnson and Peter Bobbi for technical assistance, Professor L. Mander (ANU, Canberra) and Professor B.O. Phinney (UCLA, USA) for labelled GA standards, and the Australian Research Council for financial support.     

Sequence and structural homology among membrane-associated domains of CFTR and certain transporter proteins

A sequence comparison of the two membrane-associated (MA) domains of the cystic fibrosis transmembrane conductance regulator (CFTR), multidrug resistance transporter (MDR), and -factor pheromone export system (STE6) proteins, each of which are believed to contain a total of 12 transmembrane (TM) segments, reveals significant amino acid homology and length conservation in the loop regions that connect individual TM sequences. Similar structural homology is observed between these proteins, hemolysin B (HLYB) and the major histocompatibility-linked peptide transporter, HAM1, the latter two which contain a single MA domain composed of six TM segments. In addition, there are specific sequences that are conserved within the TM segments of the five different membrane proteins. This observation suggests that the folding topologies of the MA domains of MDR, STE6, and CFTR in the plasma membrane are likely to be very similar. The sequence analysis also reveals that there are three characteristic motifs (a pair of aromatic residues, LTLXXXXXXP and GXXL) that are conserved in MDR, STE6, HLYB, HAM1, but not in CFTR. We propose that although CFTR may be evolutionarily related to these other membrane proteins, it belongs to a separate subclass.     

Evidence for the presence of a receptor for the cytolethal distending toxin (CLDT) of Campylobacter jejuni on CHO and HeLa cell membranes and development of a receptor-based enzyme-linked immunosorbent assay for detection of CLDT

Abstract Using ligand blotting, it was found that partially purified cytolethal distending toxin prepared from and enterotoxigenic strain of Campylobacter jejuni , bound to two peptides of molecular masses of approximately 59 kDa and 45 kDa and to a single peptide of 59 kDa in protein blots prepared from HeLa and CHO cell membranes, respectively. In contrast, labile toxin of Escherichia coli and cholera toxin bound to a single peptide of the same molecular mass (15 kDa) on protein blots prepared from both CHO and HeLa cell crude membranes resolved by gel electrophoresis. This banding pattern was identical using SDS-solubilized membrane, with or without heat treatment, but no band was obtained when reduced (treatment with 2-mercaptoethanol) samples were used for the gel electrophoresis. The differences between receptors of cytolethal distending toxin and cholera toxin/labile toxin were exploited to develop a receptor-based enzyme-linked immunosorbent assay for detection of cytolethal distending toxin which involved the consecutive addition of either solubilized CHO or HeLa membranes, antigen and antibody. This enzyme-linked immunosorbent assay consistently detected crude cytolethal distending toxin diluted up to 16-fold. The receptor-based enzyme-linked immunosorbent assay for detection of cytolethal distending toxin developed in this study is a suitable alternative assay which can be performed easily in laboratories with minimal facilities and, more importantly, the results are available within a few hours as compared to times of up to 5 days in the conventional tissue culture detection of cytolethal distending toxin.     

Dietary exposure and risk assessment to trace elements and iodine in seaweeds

IntroductionSeaweeds are a rich source of elements such as iodine, and are also able to accumulate contaminants such as trace elements.MethodsThe aim of this study was to assess the dietary exposure as well as the risk from iodine and trace elements in edible seaweeds for the French population using current consumption data. The contribution of seaweeds to overall dietary exposure to trace elements and iodine was evaluated, and for those substances with minimal contribution to overall dietary exposure, simulations were performed to propose increased maximal limits in seaweeds.ResultsCadmium, inorganic arsenic and mercury in seaweeds were very low contributors to total dietary exposure to these contaminants (0.7 % 1.1 % and 0.1 % on average, respectively). Dietary exposure to lead via seaweed may contribute up to 3.1 % of total dietary exposure. Dietary consumption of iodine via seaweed may contribute up to 33 % of total exposure to iodine, which makes seaweeds the strongest contributor to iodine in diet.DiscussionNew maximal values in seaweeds are proposed for the very low contributors to total dietary exposure: 1 mg/kg dw for cadmium, 10 mg/kg dw for inorganic arsenic and 0.3 mg/kg dw for mercury.     

Protein structure dynamics by crosslinking mass spectrometry

Crosslinking mass spectrometry captures protein structures in solution. The crosslinks reveal spatial proximities as distance restraints, but do not easily reveal which of these restraints derive from the same protein conformation. This superposition can be reduced by photo-crosslinking, and adding information from protein structure models, or quantitative crosslinking reveals conformation-specific crosslinks. As a consequence, crosslinking MS has proven useful already in the context of multiple dynamic protein systems. We foresee a breakthrough in the resolution and scale of studying protein dynamics when crosslinks are used to guide deep-learning-based protein modelling. Advances in crosslinking MS, such as photoactivatable crosslinking and in-situ crosslinking, will then reveal protein conformation dynamics in the cellular context, at a pseudo-atomic resolution, and plausibly in a time-resolved manner.