Stable isotope labeling of Arabidopsis thaliana for an NMR-based metabolomics approach

Nuclear magnetic resonance (NMR) will become a key technology in plant metabolomics with the use of stable isotope labeling and advanced hetero-nuclear NMR methodologies. To demonstrate the power of this approach, we performed multi-dimensional hetero-nuclear NMR analysis of metabolic movement of carbon and nitrogen nuclei in Arabidopsis thaliana. First, distinct ethanol-stress response was investigated using (13)C-labeled wild type and an ethanol-hypersensitive mutant plants. Furthermore, we followed nitrogen fluxes in (15)N-labeled seeds during the initiation of germination in vivo. The future role of stable isotope-labeling combined with advanced hetero-nuclear NMR in plant metabolomics is discussed.     

Effects of dietary supplements on space radiation-induced oxidative stress in Sprague-Dawley rats

Of particular concern for the health of astronauts during space travel is radiation from protons and high-mass, high-atomic-number (Z), and high-energy particles (HZE particles). Space radiation is known to induce oxidative stress in astronauts after extended space flight. In the present study, the total antioxidant status was used as a biomarker to evaluate oxidative stress induced by gamma rays, protons and HZE-particle radiation. The results demonstrate that the plasma level of total antioxidants in Sprague-Dawley rats was significantly decreased (P < 0.01) in a dose-dependent manner within 4 h after exposure to gamma rays. Exposure to protons and HZE-particle radiation also significantly decreased the serum or plasma level of total antioxidants in the irradiated animals. Diet supplementation with L-selenomethionine alone or a combination of selected antioxidant agents was shown to partially or completely prevent the decrease in the serum or plasma levels of total antioxidants in animals exposed to gamma rays, protons or HZE particles. These findings suggest that exposure to space radiation may compromise the capacity of the host antioxidant defense and that this adverse biological effect can be prevented at least partially by dietary supplementation with L-selenomethionine and antioxidants.     

Five new C21 steroidal glycosides from Cynanchum komarovii Al.Iljinski

Five new C21 steroidal glycosides, namely, komarosides D (1), E (2), F (3), G (4), and H (5), along with two known C21 steroidal glycosides cynatratoside E (6) and hancoside A (7), were isolated from the ethanol extract of the roots of Cynanchum komarovii Al.Iljinski (Asclepiadaceae). Their structures were determined by physiochemical and spectroscopic analysis. Among these glycosides, five had an aberrant 13,14:14,15-disecopregnane-type skeleton, and the other two had normal four-ring C21 steroid skeletons. The existence of more than one type of C21 steroid skeleton in one species is rare in the plants of the family Asclepiadaceae, and this has chemotaxonomic significance for this species.     

Marker-assisted selection in segregating generations of self-fertilizing crops

Computer simulations were used to study the efficiency of MAS for breeding self-fertilizing crops, based on a general model including additive, dominance and epistasis. It was shown that MAS not only gave larger genetic responses but also dramatically increased the frequencies of superior genotypes as compared with phenotypic selection. However, the advantages of MAS over phenotypic selection were considerably reduced when conducting selection in later generations. A modified method combining MAS in early generations with phenotypic selection in later generations was thus proposed from an efficiency standpoint. We also proposed a potential index to measure the probability of an individual showing superior genotypes under selfing. It was apparent that more superior genotypes could be derived from selection by using the potential index than by using other methods. The implications of these findings for plant breeding are discussed.Communicated by H.C. Becker     

Independent expression of the two paralogous<Emphasis Type="Italic"> CCL4</Emphasis> genes in monocytes and B lymphocytes

The CCL4 chemokine is secreted by a variety of cells following stimulation. CCL4 affects several different types of cells that are important for acute inflammatory responses and are critical for the development of specific immune responses to foreign antigens. The human genome contains two genes for the CCL4 chemokine. Although highly homologous, the two genes encode slightly different proteins. We analyzed the mRNA expressed in monocytes and B lymphocytes and found that while monocytes express predominantly one CCL4 gene, known as ACT-2, peripheral blood B lymphocytes express a mixture of ACT-2 and the second CCL4 gene, lymphocyte activating gene-1 (LAG-1). Although peripheral blood B cells, CD27^– B cells, and CD27⁺ B cells all express a mixture of LAG-1 and ACT-2, the B-cell lines that were studied regulate the two genes independently. RL, SU-DHL-6, and REH cells predominantly express LAG-1. These studies demonstrate that monocytes and B cells utilize different mechanisms to regulate expression of the two CCL4 genes and suggest that the two genes may not have identical activities.     

Two morphological types of cell appendages on a strongly adhesive bacterium, Acinetobacter sp. strain Tol 5

Two morphological types of appendages, an anchor-like appendage and a peritrichate fibril-type appendage, have been observed on cells of an adhesive bacterium, Acinetobacter sp. strain Tol 5, by use of recently developed electron microscopic techniques. The anchor extends straight to the substratum without branching and tethers the cell body at its end at distances of several hundred nanometers, whereas the peritrichate fibril attaches to the substratum in multiple places, fixing the cell at much shorter distances.     

Studies on activities, cell uptake and DNA binding of four trans-planaramineplatinum(II) complexes of the form: trans-PtL(NH3)Cl2, where L=2-hydroxypyridine, imidazole, 3-hydroxypyridine and imidazo(1,2-alpha)pyridine

Four trans-planaramineplatinum(II) complexes code named YH9, YH10, YH11 and YH12 each of the form trans-PtL(NH(3))Cl(2), where L=2-hydroxypyridine and 3-hydroxypyridine, imidazole, and imidazo(1,2-alpha)pyridine for YH9, YH10, YH11 and YH12, respectively, have been synthesized and the activity of the compounds against human cancer cell lines, cell uptake, DNA-binding and nature of interaction with pBR322 plasmid DNA have been studied. The compound having imidazo(1,2-alpha)pyridine ligand as one the carrier ligands in the trans-configuration is found to be significantly more active than cis-platin against ovarian A2780(cisR) cancer cell line corresponding with higher Pt-DNA binding. All other compounds have resistance factors less than that for cis-platin in the A2780 and A2780(cisR) cell lines. A greater prevention of BamH1 digestion with increasing concentration of the compounds indicates that as the compounds bind with nucleobases in DNA, the DNA conformation is changed sufficiently so as to prevent BamH1 digestion at the specific GG site. Gel electrophoresis results also indicate that as the compounds bind to DNA, unwinding of supercoiled form I DNA takes place to change it from the negatively supercoiled form I through relaxed circular form I to the positively supercoiled form I.     

Novel polynuclear platinum adducts detected during the reactions of [Pt(Met-S,N)Cl2] with gamma-glutathione and L-cysteine

The reactions of platinum(II) complexes with thiol containing molecules are highly relevant to the mechanism of action of platinum-based drugs. This work presents the electrospray mass spectrometry (ESMS) and NMR results on the reactions of [Pt(l-MetH-S,N)Cl(2)] (l-MetH: l-methionine) with gamma-glutathione (GSH) and l-cysteine (l-Cys) at different pH and different molar ratios. Polymeric species such as [Pt(2)(micro-SG-S)(2)(Met-S,N)(2)], [Pt(3)(micro-SG-S)(4)(Met-S,N)(2)], [Pt(4)(micro-SG-S)(6)(Met-S,N)(2)] and [Pt(5)(micro-SG-S)(8)(Met-S,N)(2)] (l-Met: deprotonated l-methionine) were detected and were stable for long hours. For both reactions, the polymerization extent decreased with the increase of pH. For the reaction of l-Cys, only mononuclear complex [Pt(l-Met-S,N)(l-Cys-S,N)] was observed when pH>9. The observation and identification of polymeric (higher than binuclear) adducts of Pt(II)/GSH and Pt(II)/l-Cys appears to be unprecedented.     

Structure,function and assembly of Photosystem II and its light-harvesting proteins

Photosystem II (PSII) is a multisubunit chlorophyll–protein complex that drives electron transfer from water to plastoquinone using energy derived from light. In green plants, the native form of PSII is surrounded by the light-harvesting complex (LHCII complex) and thus it is called the PSII–LHCII supercomplex. Over the past several years, understanding of the structure, function, and assembly of PSII and LHCII complexes has increased considerably. The unicellular green alga Chlamydomonas reinhardtii has been an excellent model organism to study PSII and LHCII complexes, because this organism grows heterotrophically and photoautotrophically and it is amenable to biochemical, genetic, molecular biological and recombinant DNA methodology. Here, the genes encoding and regulating components of the C. reinhardtii PSII–LHCII supercomplex have been thoroughly catalogued: they include 15 chloroplast and 20 nuclear structural genes as well as 13 nuclear genes coding for regulatory factors. This review discusses these molecular genetic data and presents an overview of the structure, function and assembly of PSII and LHCII complexes.     

Effect of pituitary adenylate cyclase-activating polypeptide 38 (PACAP38) on tissue oxygen content--treatment in central nervous system of mice

It has been reported that pituitary adenylate cyclase-activating polypeptide (PACAP) plays an important role in preventing neuronal cell death and is also a potent vasodilator. Cerebral hypotension and hypoperfusion during cerebral ischemia and neurodegenerative diseases are well known as some of the negative factors which aggravate neuronal cell death. Nevertheless, the effect of PACAP on the cerebral circulation was not understood well. Therefore, in the present study, we determined the mean arterial blood pressure (MBP), regional cerebral blood flow (rCBF) and cerebral oxygen content (pO2) in mice, and estimated the therapeutically useful doses of PACAP. Under barbiturate anesthesia, polyethylene tubes were inserted into mice to monitor MBP and to administer PACAP (5 x 10(-13)-5 x 10(-8) mol/kg) or vasoactive intestinal peptide (VIP; 5 x 10(-12) and 5 x 10(-9) mol/kg). Then, MBP, rCBF and cerebral pO2 were simultaneously measured in the mice. PACAP (5 x 10(-10)-5 x 10(-9) mol/kg) injections transiently decreased MBP, and cerebral pO2. PACAP (5 x 10(-8) mol/kg) injections produced a long-lasting potent decline of MBP, rCBF and cerebral pO2. Therefore, PACAP should be applied at low doses which do not influence the MBP and cerebral circulation to determine the therapeutically useful doses of PACAP for neuroprotection.