Partial decline of Arachis hypogaea L. photosynthesis triggered by drought stress

Photosynthetic capacity (PC) of three peanut cultivars (Arachis hypogaea L. cvs. 57-422, 73-30, and GC 8-35) decreased during drought stress (decline in relative water content from ca. 95 to 70 %) and recovered two days after rewatering. Mild water stress was not limiting for the total ribulose-1,5-bisphosphate carboxylase/oxygenase activity, since this enzyme activity increased under drought. Photosystem (PS) 2 and PS1 (the latter only in cv. GC 8-35) electron transport activities decreased under drought. The ratio of the variable to maximal chlorophyll fluorescence (Fv/Fm) decreased mainly in the cv. GC 8-35. All cultivars showed decreases in photochemical quenching (qP) and quantum yield of PS2 electron transport (Φe). Increase of basal fluorescence (F0) was observed in the cvs. 73-30 and GC 8-35, while the cv 57-422 showed a decrease. After rewatering a sharp increase was observed in the majority of the parameters. Thus under the present stress conditions, the cv GC 8-35 was the most affected for all the parameters under study. The cv. 57-422 showed a higher degree of tolerance being gradually affected in photosynthetic capacity (PC) in contrast to the two other cvs. which showed a sharp decrease in PC at the beginning of the drought cycle. This revised version was published online in September 2006 with corrections to the Cover Date.     

Small-molecule active pharmaceutical ingredients of approved cancer therapeutics inhibit human aspartate/asparagine-β-hydroxylase

Human aspartate/asparagine-β-hydroxylase (AspH) is a 2-oxoglutarate (2OG) dependent oxygenase that catalyses the hydroxylation of Asp/Asn-residues of epidermal growth factor-like domains (EGFDs). AspH is reported to be upregulated on the cell surface of invasive cancer cells in a manner distinguishing healthy from cancer cells. We report studies on the effect of small-molecule active pharmaceutical ingredients (APIs) of human cancer therapeutics on the catalytic activity of AspH using a high-throughput mass spectrometry (MS)-based inhibition assay. Human B-cell lymphoma-2 (Bcl-2)-protein inhibitors, including the (R)-enantiomer of the natural product gossypol, were observed to efficiently inhibit AspH, as does the antitumor antibiotic bleomycin A₂. The results may help in the design of AspH inhibitors with the potential of increased selectivity compared to the previously identified Fe(II)-chelating or 2OG-competitive inhibitors. With regard to the clinical use of bleomycin A₂ and of the Bcl-2 inhibitor venetoclax, the results suggest that possible side-effects mediated through the inhibition of AspH and other 2OG oxygenases should be considered.     

A Single Nucleotide Polymorphism of Chicken Acetyl-CoA Carboxylase A Gene Associated with Fatness Traits

Acetyl-CoA carboxylase α (ACCα) is a major rate-limiting enzyme in the biogenesis of long-chain fatty acids. It can catalyze the carboxylation of acetyl-CoA to form malonyl-CoA that plays a key role in the regulation of fatty acid metabolism. The objective of the present study was to investigate the associations of ACCα gene polymorphisms with chicken growth and body composition traits. The Northeast Agricultural University broiler lines divergently selected for abdominal fat content and the Northeast Agricultural University F₂ Resource Population were used in the current study. Body weight and body composition traits were measured in the aforementioned two populations. A synonymous mutation was detected in the exon 19 region of ACCα gene, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was developed to genotype all the individuals derived from the aforementioned populations. Association analysis revealed that the polymorphism was associated with abdominal fat weight and percentage of abdominal fat in the two populations. The results suggested that ACCα gene could be a candidate locus or linked to a major gene that affects abdominal fat content in the chicken.     

Redox regulation and oxidant activation of heme oxygenase-1

The ultraviolet A (UVA, 320–400 nm) component of sunlight has the potential to generate an oxidative stress in cells and tissue so that antioxidants (both endogenous and exogenous) strongly influence the biological effects of UVA. The expression of several genes (including heme oxygenase-1, HO-1; collagenase; the CL100 phosphatase and the nuclear oncogenes, c-fos and c-jun) is induced following physiological doses of UVA to cells and this effect can be strongly enhanced by removing intracellular glutathione or enhancing singlet oxygen lifetime. We have observed that heme is released from microsomal heme-containing proteins by UVA and other oxidants and that activation of HO-1 expression by UVA correlates with levels of heme release. UVA radiation also leads to an increase in labile iron pools (either directly or via HO-1) and eventual increases in ferritin levels. The role of heme oxygenase in protection of skin fibroblasts is probably an emergency inducible defense pathway to remove heme liberated by oxidants. The slower increase in ferritin levels is an adaptive response which serves to keep labile iron pools low and thereby reduce Fenton chemistry and oxidant-induced chain reactions involving lipid peroxidation. In keratinocytes, the primary target of UVA radiation, heme oxygenase levels are constitutively high (because of HO-2 expression). Since there is a corresponding increase in basal levels of ferritin the epidermis appears to be well protected constitutively against the oxidative stress generated by UVA.     

The Baseline Cosinus Function: A Periodic Regression Model for Biological Rhythms

Abstract

Both tryptophan oxygenase and tyrosine aminotransferase of rat liver show diurnal variations in the inducibility by quinolinic acid. The maxima of effectiveness of quinolinic acid precede the maxima of normal enzyme activity. In the case of tyrosine aminotransferase, the induction kinetics and the dose response curve were also greatly depending on the time of day. No rhythmicity could be detected in the activities of 3‐hydroxyanthranilate oxygenase and ornithine aminotransferase.     

Screening and development of enzymes for determination and transformation of amino acids

ABSTRACT

The high stereo- and substrate specificities of enzymes have been utilized for micro-determination of amino acids. Here, I review the discovery of l-Phe dehydrogenase and its practical use in the diagnosis of phenylketonuria in more than 5,400,000 neonates over two decades in Japan. Screening and uses of other selective enzymes for micro-determination of amino acids have also been discussed. In addition, novel enzymatic assays with the systematic use of known enzymes, including assays based on a pyrophosphate detection system using pyrophosphate dikinase for a variety of l-amino acids with amino-acyl-tRNA synthetase have been reviewed. Finally, I review the substrate specificities of a few amino acid-metabolizing enzymes that have been altered, using protein engineering techniques, mainly for production of useful chemicals, thus enabling the wider use of natural enzymes.     

Biomarkers of exposure in fish inhabiting the Swan–Canning Estuary, Western Australia – a preliminary study

The estuarine portion of the Swan–Canning riversystem runs through the centre of Perth,Western Australia's capital city, with apopulation of approximately 1.4 million people. Little is known about impact of chemicalsentering the estuary via road runoff andstormwater drains on biota inhabiting thesystem. Black bream (Acanthopagrusbutcheri) were collected from seven sites inthe Swan–Canning estuary during August andSeptember 2000, at the end of the winter (wet)season. Serum sorbitol dehydrogenase (s-SDH)was unaffected by the sex of the fish and nosignificant differences were observed betweenthe sites indicating that the measuredethoxyresorufin-O-deethylase (EROD)activity was not hindered by hepatic tissuedamage. The black bream were in an advancedstage of gonad maturation, which affected ERODhepatic activity with lower EROD activity infemale compared to male fish. EROD activityand bile metabolite levels were significantlyhigher at the site closest to the Perth CentralBusiness District, while most downstream sitewas the least impacted, which may be due totidal flushing of the lower estuary by marinewaters. The ratio of naphthalene-type tobenzo(a) pyrene (B(a)P)-typemetabolites suggests that the source ofpetroleum hydrocarbons within the river systemis a mixture polycyclic aromatic hydrocarbons(PAHs) from pyrolytic origin and from unburntfuels. Biomarker levels in the black breamindicate that major roads and drains aresignificant contributors of mixed functionoxygenase (MFO) inducing chemicals includingpolycyclic aromatic hydrocarbons (PAH) into theSwan–Canning estuary and that there is noupstream or downstream gradient in biomarkerresponse.     

Suitable conditions for characterization,identification, and isolation of the mRNA of the small subunit of ribulose-1,5-bisphosphate carboxylase from Nicotiana sylvestris

The products synthesized in vitro by messenger RNA (mRNA) extracted from Nicotiana sylvestris were analyzed by electrophoresis on polyacrylamide slab gels. Only three of the major polypeptides synthesized are considered here: P₅₅, P₃₂, and P₂₀. P₅₅ and P₃₂ were translated from chloroplast mRNA. P₅₅ corresponds to the large subunit of ribulose-1,5-bisphosphate (RuP₂) carboxylase; P₃₂ is probably a chloroplast membrane protein. P₂₀, the polypeptide synthesized from cytoplasmic poly(A)⁺ RNA, is the precursor of the small subunit of RuP₂ carboxylase. The balance between P₂₀ and P₃₂, in which their relative proportions varied inversely, was regulated by the age of the leaves and the time of illumination; we took advantage of this phenomenon to isolate the mRNA from the small subunit in relatively large amounts. This mRNA has a molecular weight of 350,000.Abbreviations RuP₂ ribulose-1,5-bisphosphate - mRNA messenger RNA - SDS sodium dodecyl sulfate     

Analysis of gene expression profile in p130(Cas)-deficient fibroblasts

p130(Cas) (Cas) is a docking protein that becomes tyrosine phosphorylated in v-Src- or v-Crk-transformed cells and in integrin-stimulated cells. Cas -/- fibroblasts show defects in stress fiber formation, cell spreading, cell migration, and transformation by activated Src. To further characterize the role of Cas in signaling, we compared the expression profile in Cas -/- fibroblasts with that in Cas-re-expressing fibroblasts using the microarray methods. In Cas -/- fibroblasts, the expression of heme oxygenase 1 and caveolin-1 was reduced, but the expression of procollagen 1 alpha 1, procollagen 3 alpha 1, procollagen 11 alpha 1, elastin, periostin, TSC-36, and MARCKS was enhanced. The domains in Cas necessary for the change varied among these genes. Activated Src reduced the expression of most of these genes both in Cas -/- and in Cas +/+ fibroblasts. These results suggest the existence of signaling pathways that emanate from Cas to gene expression.     

Absence of multiple forms of ribulose 15-bisphosphate carboxylase/oxygenase in mung bean

Ribulose 1,5-bisphosphate carboxylase/oxygenase has been reported to occur in multiple forms in mung bean (Phaseolus aureus) using Sephadex G-200 chromatography. We have isolated this enzyme by identical methodology. The profile from Sephadex G-200 chromatography shows only one peak in contrast to the previous report and we find no evidence to corroborate the conclusions. Where V_c, V_o and K_c, K_o represent V_max and Michaelis constants, respectively, the constant V_cK_o/V_oK_c for the single form is 70 at 40 μM CO₂ and 1200 μM O₂.