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Query: UNIPROT:P06889 (Mol)
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We examined biochemical changes accompanying feeding and starvation from hatch to Stage VI (day 74 after hatch) in spiny lobster, Jasus edwardsii, phyllosoma larvae. Larval dry weights (dw) increased 17-fold from hatch (80+/-1 microg) to Stage VI (1415+/-44 microg). Larvae starved for 6-11 days at Stages II, IV and VI were 14-40% lighter than their fed counterparts fed enriched Artemia. The increases and losses in total dry weight during feeding and starvation were associated with changes in the content of protein (constituting 31.4-41.7% of dw) and carbohydrate (constituting 2.6-5.3% of dw), while larger changes in lipid content indicated its greater importance as an energy substrate. Lipid content increased from 7.9% of dw at hatch to its highest of 12.5% at Stage IV, but declined by 50% or more during starvation. This suggests that protein, carbohydrate and lipid are all important energy stores, although lipids are catabolized at a greater rate during food deprivation. The principal lipid class was polar lipid (PL; 79-92% of total lipid), followed by sterol (ST; 6-20%), with triacylglycerol and other lipid classes at <2%. PL were catabolized and ST were conserved during starvation. Changes in the fatty acid (FA) profile had mostly occurred before the first moult at day 8 after hatch, with gradual changes thereafter to Stage VI, reflecting their abundance in the Artemia diet. There was some conservation of the major essential FAs, 20:4n-6, 20:5n-3, 22:6n-3, and the FA profile showed large gains in the C(18) polyunsaturated FA, 18:1n-9, 18:2n-6. Ascorbic acid content increased 10-fold from hatch to the end of Stage I (36 and 333 microgg(-1) dw, respectively), while the content at the end of Stage II was higher in fed than that in starved larvae (439 and 174 microgg(-1) dw, respectively). Our study will assist in the development of alternatives to nutritionally incomplete diets, such as live ongrown Artemia, to meet the requirements of phyllosoma in culture.
Comp Biochem Physiol A Mol Integr Physiol 2003 Oct
PMID:Biochemical composition during growth and starvation of early larval stages of cultured spiny lobster (Jasus edwardsii) phyllosoma. 1451 54

Vitamin C is transported as ascorbic acid (AA) through the sodium-ascorbate cotransporters (SVCT1 and -2) and as dehydroascorbic acid (DHA) through the facilitative glucose transporters. All cells have glucose transporters and take up DHA that is trapped intracellularly by reduction and accumulated as AA. SVCT2 is widely expressed in cells and tissues at the mRNA level; however, only specialized cells directly transport AA. We undertook a molecular analysis of SVCT2 expression and discovered a transcript encoding a short form of human SVCT2 (hSVCT2-short) in which 345 bp is deleted without a frame shift. The deletion involves domains 5 and 6 and part of domain 4. cDNA encoding this isoform was isolated and expressed in 293T cells, where the protein was detected on the plasma membrane. Transport studies, however, revealed that hSVCT2-short gave rise to a nonfunctional transporter protein. hSVCT2-short arises by alternative splicing and encodes a protein that strongly inhibited the function of SVCT2 and, to a lesser extent, SVCT1 in a dominant-negative manner, probably by protein-protein interaction. The expression of hSVCT2-short varies among cells. PCR analysis of cDNA isolated from melanocytes capable of transporting AA revealed a predominance of the full-length isoform, while HL-60 cells, which express SVCT2 at the mRNA level and were incapable of transporting AA, showed a predominance of the short isoform. These findings suggest a mechanism of AA uptake regulation whereby an alternative SVCT2 gene product inhibits transport through the two known AA transporters.
Mol Cell Biol 2004 Apr
PMID:A human sodium-dependent vitamin C transporter 2 isoform acts as a dominant-negative inhibitor of ascorbic acid transport. 1506 Jan 39

Vitamin C (L-ascorbic acid) has important antioxidant and metabolic functions in both plants and animals, humans have lost the ability to synthesize it. Fresh produce is the major source of vitamin C in the human diet yet only limited information is available concerning its route(s) of synthesis in plants. In contrast, the animal vitamin C biosynthetic pathway has been elucidated since the 1960s. Two biosynthetic pathways for vitamin C in plants are presently known. The D-mannose pathway appears to be predominant in leaf tissue, but a D-galacturonic acid pathway operates in developing fruits. Our group has previously shown that transforming lettuce and tobacco with a cDNA encoding the terminal enzyme of the animal pathway, L-gulono-1,4-lactone oxidase (GLOase, EC 1.1.3.8), increased the vitamin C leaf content between 4- and 7-fold. Additionally, we found that wild-type (wt) tobacco plants had elevated vitamin C levels when fed L-gulono-1,4-lactone, the animal precursor. These data suggest that at least part of the animal pathway may be present in plants. To further investigate this possibility, wild-type and vitamin-C-deficient Arabidopsis thaliana (L.) Heynh (vtc) plants were transformed with a 35S: GLOase construct, homozygous lines were developed, and vitamin C levels were compared to those in untransformed controls. Wild-type plants transformed with the construct showed up to a 2-fold increase in vitamin C leaf content compared to controls. All five vtc mutant lines expressing GLOase had a rescued vitamin C leaf content equal or higher (up to 3-fold) than wt leaves. These data and the current knowledge about the identity of genes mutated in the vtc lines suggest that an alternative pathway is present in plants, which can bypass the deficiency of GDP-mannose production of the vtc1-1 mutant and possibly circumvent other steps in the D-mannose pathway to synthesize vitamin C.
Plant Mol Biol 2003 Dec
PMID:L-Gulono-1,4-lactone oxidase expression rescues vitamin C-deficient Arabidopsis (vtc) mutants. 1508 29

Unsaturated lipids in sperm plasma membranes are very susceptible to peroxidation when exposed to reactive oxygen species (ROS). In this investigation we have incubated ram spermatozoa in the presence of two ROS generating systems, ascorbate/FeSO4 and potassium peroxychromate (K3CrO8), and examined their effects on membrane fluidity by measuring fluorescence recovery after photobleaching (FRAP) of a lipid reporter probe 5-(N-octadecanoyl)-aminofluorescein (ODAF). Peroxidation was monitored by malonaldehyde formation and changes in fluorescence emission of 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C11-BODIPY(581/591)). Ascorbate/FeSO4-induced peroxidation was inhibited by Vitamin E, butylated hydroxyanisole (BHA), 1,4-diazobicyclo(2,2,2)octane (DABCO), and to a lesser extent by ethanol. Added superoxide dismutase (SOD), gluthathione peroxidase (GPX), and catalase were ineffective scavengers. K3CrO8 induced very rapid peroxidation that could be delayed, but not prevented, by Vitamin E, BHT, DABCO, ethanol, and mannitol; once again SOD, GPX, and catalase were ineffective scavengers. Neither peroxidation with ascorbate/FeSO4 nor K3CrO8, or added H2O2 or malonaldehyde perturbed ODAF diffusion in any region of the sperm plasma membrane. Vitamin E tended to enhance diffusion rates. Exogenous cumene hydroperoxide, however, reduced ODAF diffusion to low levels on the sperm head. These results suggest that the adverse effects of ROS on spermatozoa are more likely to be caused by direct oxidation of proteins and membrane permeabilisation than disturbance of lipid fluidity.
Mol Reprod Dev 2004 Jul
PMID:Lipid diffusion in sperm plasma membranes exposed to peroxidative injury from oxygen free radicals. 1511 31

Lateral gene transfer has emerged as an important force in bacterial evolution. A substantial number of genes can be inserted into or deleted from genomes through the process of lateral transfer. In this study, we looked for atypical occurrence of genes among related organisms to detect laterally transferred genes. We have analyzed 50 bacterial complete genomes from nine groups. For each group we use a 16s rRNA phylogeny and a comparison of protein similarity to map gene insertions/deletions onto their species phylogeny. The results reveal that there is poor correlation of genes inserted, deleted, and duplicated with evolutionary branch length. In addition, the numbers of genes inserted, deleted, or duplicated within the same branch are not always correlated with each other. Nor is there any similarity within groups. For example, in the Rhizobiales group, the ratio of insertions to deletions in the evolutionary branch leading to Agrobacterium tumefaciens str. C58 (Cereon) is 0.52, but it is 39.52 for Mesorhizobium loti. Most strikingly, the number of insertions of foreign genes is much larger in the external branches of the trees. These insertions also greatly outnumber the occurrence of deletions, and yet the genome sizes of these bacteria remain roughly constant. This indicates that many of the insertions are specific to each organism and are lost before related species can evolve. Simulations of the process of insertion and deletion, tailored to each phylogeny, support this conclusion.
Mol Biol Evol 2004 Jul
PMID:Patterns of bacterial gene movement. 1511 2

The absorption spectral change of methyl glyoxal (MG) due to the interaction with ascorbic acid (AA or Vitamin C) has been investigated using steady-state spectroscopic technique. A plausible explanation for the spectral change has been discussed on the basis of hydrogen bonding interaction between the two interacting species. The equilibrium constant for the complex formation due to hydrogen bonding interaction between MG and AA has been obtained from absorption spectral changes. Ab inito calculations with DFT B3LYP/6/31G (d,p) basis sets have been used to find out the molecular structure of the hydrogen bonded complex. The O...H distance found in the O-H...O hydrogen bond turns out to be quite short (1.974 A) which is in conformity with the large value of the equilibrium constant determined experimentally.
Spectrochim Acta A Mol Biomol Spectrosc 2004 Jun
PMID:Interaction between methyl glyoxal and ascorbic acid: experimental and theoretical aspects. 1514 93

Cadmium (Cd) is one of the environmental pollutants affecting various tissues and organs including testis. Harmful effect of Cd in testis is known to be germ cell degeneration and impairment of testicular steroidogenesis. Animals treated with high doses of Cd (0.2 and 0.3 mg/100g BW) showed a significant decrease in serum testosterone (T) level, but a significant induction of testicular lipid peroxidation levels. TUNEL assay showed that low doses of Cd (0.13 and 0.15 mg/100g BW) exhibited typical characteristics of apoptosis while high doses of Cd caused more necrosis than apoptosis. In contrast, supplementation with ascorbic acid reduced testicular lipid peroxidation levels. Ascorbic acid supplementation restored testicular 3beta-hydroxysteroiddehydrogenase (HSD) and 17beta-HSD enzyme activities, 3beta-HSD and cytochrome P450 side chain cleavage (P450(scc)) mRNA levels and serum T concentration to normal in Cd-administered rats. Moreover, administration of ascorbic acid prevented germ cell apoptosis as demonstrated by the reduced number of TUNEL-positive cells in germinal epithelium and inhibited Cd-induced necrosis. These results indicate that ascorbic acid have protective roles in vivo on the Cd-induced overall testicular damage including impaired steroidogenesis and germ cell death possibly through scavenging the reactive oxygen species generated by Cd administration.
Mol Cell Endocrinol 2004 Jun 30
PMID:Effect of ascorbic acid supplementation on testicular steroidogenesis and germ cell death in cadmium-treated male rats. 1522 32

New organotin(IV) ascorbates of the general formulae R(3)Sn(HAsc) (where R = Me , n-Pr, n-Bu and Ph) and R(2)Sn(Asc) (where R = n-Bu and Ph) have been synthesized by the reaction of R(n)SnCl(4-n) (where n = 2 or 3) with monosodium-l-ascorbate. The bonding and coordination behaviour in these complexes are discussed on the basis of UV-Vis, IR, Far-IR, (1)H and (13)C NMR, and (119)Sn Mossbauer spectroscopic studies. L-Ascorbic acid acts as a monoanionic bidentate ligand in R(3)Sn(HAsc) coordinating through O(1) and O(3). The Mossbauer studies together with IR and NMR studies suggest that for these polymeric derivatives, the polyhedron is trigonal bipyramidal around tin with three organic groups in the equatorial positions. In R(2)Sn(Asc), L-ascorbic acid acts as dianionic tetradentate ligand and a polymeric structure with octahedral geometry around tin with trans organic groups has been tentatively proposed. The complexes have been assayed for their anti-inflammatory and cardiovascular activity. Ph(2)Sn(Asc) has been found to show the highest activity among the studied complexes. It is suggested on the basis of potentiometric studies of Me(2)Sn(IV) and Me(3)Sn(IV) systems with L-ascorbic acid that under physiological conditions (pH = 7.0) Me(2)Sn(HAsc)(OH) (approximately 60%), Me(2)Sn(OH)(2) (approximately 40%) and Me(3)Sn(HAsc) (approximately 60%), Me(3)Sn(OH) (approximately 40%), respectively, are existing, which may be responsible for their biological activities.
Spectrochim Acta A Mol Biomol Spectrosc 2005 Jan 01
PMID:New organotin(IV) ascorbates: synthesis, spectral characterization, biological and potentiometric studies. 1555 24

Ascorbate peroxidase (APx) is a class I peroxidase that catalyzes the conversion of H(2)O(2) to H(2)O and O(2) using ascorbate as the specific electron donor. This enzyme has a key function in scavenging reactive oxygen species (ROS) and the protection against toxic effects of ROS in higher plants, algae, and Euglena. Here we report the identification of an APx multigene family in rice and propose a molecular evolutionary relationship between the diverse APx isoforms. In rice, the APx gene family has eight members, which encode two cytosolic, two putative peroxisomal, and four chloroplastic isoforms, respectively. Phylogenetic analyses were conducted using all APx protein sequences available in the NCBI databases. The results indicate that the different APx isoforms arose by a complex evolutionary process involving several gene duplications. The structural organization of APx genes also reflects this process and provides evidence for a close relationship among proteins located in the same subcellular compartment. A molecular evolutionary pathway, in which cytosolic and peroxisomal isoforms diverged early from chloroplastic ones, is proposed.
J Mol Evol 2004 Dec
PMID:Analysis of the molecular evolutionary history of the ascorbate peroxidase gene family: inferences from the rice genome. 1559 8

Adriamycin (ADR), a cytotoxic antineoplastic drug is used in the treatment of various solid tumors. However, its efficacy continues to be challenged by significant toxicities including testicular toxicity. In the present study, the effect of lipoic acid, a "universal antioxidant" was investigated on ADR induced peroxidative damages in rat testis. Adult male albino rats of Wistar strain were administered ADR (1 mg/kg body weight, i.v.) once a week for 10 weeks. ADR injected rats showed a significant decline in the activities of enzymic antioxidants (catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase) and non-enzymic antioxidants (reduced glutathione, Vitamin A, Vitamin C and Vitamin E) with high malondialdehyde levels. The extent of testicular toxicity was evident from the decreased activities of testicular marker enzymes (sorbitol dehydrogenase and glucose-6-phosphate dehydrogenase). Treatment with lipoic acid (35 mg/kg body weight, i.p.) one day prior to ADR administration, maintained near normal activities of the enzymes and significantly reduced lipid peroxidation, thereby proving it to be an effective cytoprotectant.
Mol Cell Biochem 2004 Dec
PMID:Remedial effect of DL-alpha-lipoic acid against adriamycin induced testicular lipid peroxidation. 1566 3


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