Examination of the pronounced increase in auxin content of senescent leaves

When a conventional IAA extraction procedure was used, the levelof auxin in senescent leaves was ten times that in young leaves.The level estimated by an improved procedure, in which IPyrAwas excluded at a high rate from the extract, was only onefourthto one-fifth and less than one-tenth that obtained by the conventionalprocedure in senescent leaves of bean and pea plants, respectively.The Trp content of detached leaves increased with the progressof leaf senescence, though TTA activity decreased. The highlevel of auxin in senescent leaves might be largely due to contaminationwith IPyrA. (Received October 30, 1978; )     

Relation of Chemotactic Response to the Amount of Receptor: Evidence for Different Efficiencies of Signal Transduction

We determined the content of galactose-glucose-, maltose-, and ribose-binding proteins in cells of Escherichia coli K-12 grown in a variety of media and also measured the respective transport and chemotactic activities that depend on those binding proteins. Correlation of the level of induction of a particular binding protein with the extent of tactic activity mediated by that protein indicates that the magnitude of the tactic response to a particular stimulating compound is a direct function of the number of receptors per cell. In contrast, comparison of the magnitudes of response to substances recognized by independent receptors indicates that some stimulus-receptor complexes are more effective in eliciting tactic responses than are others. Thus, the magnitude of response to any particular stimulating compound is a function both of the number of receptors per cell and of the effectiveness of the stimulus-receptor complex. Considerations of available information about the tactic response to maltose suggest that the effectiveness of a stimulus-receptor complex is related to the transducer with which the receptor interacts. The tar product appears to be a relatively effective transducer of the signals it accepts from receptors for aspartate, alpha-methylaspartate, and maltose, whereas the trg product appears to be a relatively ineffective transducer of signals it accepts from receptors for galactose and ribose.     

Angiogenic strategy for human ischemic heart disease: Brief overview

In the Western World ischemic coronary disease is the leading cause of morbidity and mortality. Therapeutic approaches mostly aim to restore flow to a localized segment by angioplasty or bypass surgery. Therapeutic angiogenesis and or arteriogenesis describes a strategy where blood vessel formation is induced for the purposes of treating and/or preventing ischemic disease. At present, at least 17 clinical trials of myocardial angiogenesis have been presented involving over 900 patients. Therapeutic angiogenesis makes use of the administration of angiogenic growth factor protein or gene to promote the development of endogenous collateral vessels in ischemic myocardium. Most recently, interest has grown in the potential angiogenesis effects of cell therapy—such as autologous bone marrow cells or cultured stem cells—and there are now several groups initiating phase I/II trials in this area. (Mol Cell Biochem 264: 143–149, 2004)     

Changes in structural and functional properties of oxygen-evolving complex induced by replacement of D1-glutamate 189 with glutamine in photosystem II: ligation of glutamate 189 carboxylate to the manganese cluster

A carboxylate group of D1-Glu-189 in photosystem II has been proposed to serve as a direct ligand for the manganese cluster. Here we constructed a mutant that eliminates the carboxylate by replacing D1-Glu-189 with Gln in the cyanobacterium Synechocystis sp. PCC 6803, and we examined the resulting effects on the structural and functional properties of the oxygen-evolving complex (OEC) in photosystem II. The E189Q mutant grew photoautotrophically, and isolated photosystem II core particles evolved oxygen at approximately 70% of the rate of control wild-type particles. The E189Q OEC showed typical S(2) state electron spin resonance signals, and the spin center distance between the S(2) state manganese cluster and the Y(D) (D2-Tyr-160), detected by electron-electron double resonance spectroscopy, was not affected by this mutation. However, the redox potential of the E189Q OEC was considerably lower than that of the control OEC, as revealed by the elevated peak temperature of the S(2) state thermoluminescence bands. The mutation resulted in specific changes to bands ascribed to the putative carboxylate ligands for the manganese cluster and to a few carbonyl bands in mid-frequency (1800 to 1100 cm(-1)) S(2)/S(1) Fourier transform infrared difference spectrum. Notably, the low frequency (650 to 350 cm(-1)) S(2)/S(1) Fourier transform infrared difference spectrum was also uniquely changed by this mutation in the frequencies for the manganese cluster core vibrations. These results suggested that the carboxylate group of D1-Glu-189 ligates the manganese ion, which is influenced by the redox change of the oxidizable manganese ion upon the S(1) to S(2) transition.     

Disruption of nuclear vitamin D receptor gene causes enhanced thrombogenicity in mice

Vitamin D metabolites influence the expression of various genes involved in calcium homeostasis, cell differentiation, and regulation of the immune system. Expression of these genes is mediated by the activation of the nuclear vitamin D receptor (VDR). Previous studies have shown that a hormonally active form of vitamin D, 1alpha,25-dihydroxyvitamin D3, exerts anticoagulant effects in cultured monocytic cells. To clarify whether activation of VDR plays any antithrombotic actions in vivo, hemostatic/thrombogenic systems were examined in normocalcemic VDR knock-out (KO) mice on a high calcium diet and compared with wild type and hypocalcemic VDRKO mice that were fed a regular diet. Platelet aggregation was enhanced significantly in normocalcemic VDRKO mice compared with wild type and hypocalcemic VDRKO mice. Aortic endothelial nitric-oxide (NO) synthase expression and urinary NOx excretions were reduced in hypocalcemic VDRKO mice, but not in normocalcemic VDRKO mice. Northern blot and RT-PCR analyses revealed that the gene expression of antithrombin in the liver as well as that of thrombomodulin in the aorta, liver and kidney was down-regulated in hypo- and normocalcemic VDRKO mice. Whereas tissue factor mRNA expression in the liver and kidney was up-regulated in VDRKO mice regardless of plasma calcium level. Furthermore, VDRKO mice manifested an exacerbated multi-organ thrombus formation after exogenous lipopolysaccharide injection regardless of the calcemic conditions. These results demonstrate that activation of nuclear VDR elicits antithrombotic effects in vivo, and suggest that the VDR system may play a physiological role in the maintenance of antithrombotic homeostasis.     

Interactions of Transthyretin (TTR) and Retinol-Binding Protein (RBP) in the Uptake of Retinol by Primary Rat Hepatocytes

The mechanism by which cells take up retinol from retinol-binding protein (RBP) and the role of the RBP–transthyretin (TTR) complex remain unclear. Here we report on retinol uptake through the RBP–TTR complex by primary cultured rat hepatocytes (parenchymal cells, PC) and nonparenchymal cells (NPC) following incubation with [³H]retinol–RBP or the [³H]retinol–RBP–TTR complex under several conditions. The cellular accumulation of retinol was time and temperature dependent in both PC and NPC. Analysis by HPLC showed that the incorporated [³H]retinol in NPC was mainly converted to retinyl ester, although in PC it remained mainly as unesterified retinol. However, the amount of retinol taken up from the RBP–TTR complex was nearly twofold greater than that from RBP alone. The uptake of [³H]retinol from protein-bound retinol was inhibited by an excess of either retinol–RBP or retinol–RBP–TTR complex. Moreover, retinol uptake through the RBP–TTR complex was inhibited by an excess of free TTR. From these results we postulate that TTR may take part as a positive regulator in the delivery of RBP-bound retinol from plasma, possibly by a membrane receptor, and that retinol uptake takes place preferentially from the RBP–TTR complex into both PC and NPC. The uptake of [³H]retinol (2 μM) by PC was saturated, whereas uptake by NPC was not. These results indicate that the physiological importance of TTR in retinol delivery may be especially important to vitamin A-storing stellate (Ito) cells in the NPC fraction.     

Allelic variants of the human MHC class I chain-related B gene (MICB)

 The human major histocompatibility complex (MHC) is located within a 4 megabase segment on chromosome 6p21.3. Recently, a highly divergent MHC class I chain-related gene family, MIC was identified within the class I region. The MICA and MICB genes in this family have unique patterns of tissue expression. The MICA gene is highly polymorphic, with more than 20 alleles identified to date. To elucidate the extent of MICB allelic variations, we sequenced exons 2 (α1), 3 (α2), 4 (α3), and 5 (transmembrane) as well as introns 2 and 4 of this gene in 46 HLA homozygous B-cell lines. We report the identification of eleven alleles based on seven non-synonymous, two synonymous, and four intronic nucleotide variations. Interestingly, one allele has a nonsense mutation resulting in a premature termination codon in the α2 domain. Thus, MICB appears to have fewer alleles than MICA, not unlike the allelic ratio between the HLA-C and -B loci. A preliminary linkage analysis of the MICB alleles with those of the closely located MICA and HLA-B genes revealed no conspicuous linkage disequilibrium between them, implying the presence of a potential recombination hotspot between the MICB and MICA genes. Received: 16 April 1997 / Revised: 19 May 1997