Effects of phosphorus and vitamin C deficiency,vitamin A toxicity,and lipid peroxidation on skeletal abnormalities in Atlantic halibut (Hippoglossus hippoglossus)

Dietary nutrients play an important role in skeletal tissue metabolism of fish. Deficiency and toxicity of certain nutrients have been linked to bone deformities in larval and juvenile fish. The pathogenesis of skeletal disorders in larval and juvenile fish from the same genetic stock, cultured under similar environment conditions is often difficult to distinguish when marginal deficiencies of multiple nutrients are involved. A study was conducted to characterize the skeletal deformities linked to the deficiency of phosphorus and ascorbic acid, vitamin A toxicity and lipid peroxidation in juvenile halibut. Five experimental diets containing a low level of phosphorus (0.5% dry matter basis), no vitamin C supplement, high level of vitamin A (80 000 IU kg^?1) and oxidized marine fish oil (peroxide value, 7.53 meq kg^?1) and a control diet based on cod fillet and vitamin free casein were fed to juvenile Atlantic halibut for 14 weeks in an attempt to characterize the skeletal deformities. Phosphorus, ascorbic acid, retinol, and α‐tocopherol concentrations of liver and kidney were measured at 0 and 14 weeks. Reduced vertebral ash and phosphorus content were observed in fish fed the low phosphorus diet. Skeletal abnormalities included abnormal hemal and neural spines in the hemal region and scoliosis in the cephalic and hemal regions of the vertebral column. Hepatic and kidney ascorbic acid concentrations were significantly lower in the group fed no ascorbic acid supplement. Skeletal abnormalities were scoliosis and lordosis primarily in the hemal region of the vertebral column. High levels of vitamin A in the diet caused increased hepatic retinol content and scoliosis spanning the cephalic/prehemal and anterior hemal regions of the vertebral column. Fish fed the oxidized oil diet showed increased thiobarbituric acid (TBA) value in the liver and muscle tissue with no significant decrease in hepatic vitamin E concentration. The most frequent skeletal deformity observed was scoliosis, spanning the cephalic/prehemal regions as well as the anterior hemal region of the vertebral column. The pattern and type of abnormalities observed in fish fed these experimental diets were similar to those observed in a commercial halibut hatchery.     

A histochemical study of meiocytes, microspores, pollen and the tapetum in Kalanchoe

A histochemical study was made of developing sporogenous cells, meiocytes, microspores, pollen and the tapetum in anthers of Kalanchoë morlagei. Storage polysaccharides were seen only in mature pollen. Ascorbic acid was not found in the sporogenous cells, but in meiocytes a high quantity of this compound occurred in the cytoplasm. Spore tetrads, microspores and pollen also had a high ascorbic acid content. The amounts of RNA and proteins were high in the sporogenous cells and in meiocytes during meiosis–I, but a small reduction trend with respect to RNA content was noticed. Microspores in the tetrad showed high amounts of RNA and proteins. In the young microspores RNA and proteins declined. Later, as the microspores matured, an increase in content of RNA and proteins took place. The wall of the young microspores gave a faint green colour with azure B stain, the intensity of which increased and remained high in the exine of the mature pollen. The additional wall thickening around the meiocytes and tetrads gave a strong pink colour with PAS test. This thickening showed presence of silver granules when tested for ascorbic acid, the tapetum synthesized abundant quantities of PAS positive starch, ascorbic acid, RNA and proteins from its appearance in the anther wall until microspore formation. During meiocyte meiosis the tapetum became highly vesicular. Our results indicate that the tapetum constitutes a tissue specialized for storing and supplying basic nutritive substances for the developing pollen in the anther.     

The effects of Penicillium digitatum and Fusarium oxysporum rots on nutritional content of pawpaw (Carica papaya L.)

Investigations were carried out on the effects of Penicillium digitatum and Fusarium oxysporum on the nutritional value of pawpaw (Carica papaya). Decreases were observed in ash content, phosphorus, sodium, reducing sugars and ascorbic acid levels of fruits infected with P. digitatum, but increases in calcium and potassium content. In fruits infected by F. oxysporium, there were decreases in phosphorus, calcium, sodium ascorbic acid and reducing sugar levels; but the levels of ash content increased. The total protein level increased in the fruits infected with both fungi. These results revealed a reduction in fruit quality.     

Chemical Control of Ripening in Banana

A post-harvest dip of banana (Musa paradisiaca L.) fruits into the aqueous solutions of abscisic acid and indoleacetic acid significantly hastened the banana ripening, judging from increases in total sugars, acidity, ascorbic acid and units of von Losecke's colour chart during storage at 20°C. The treatment of bananas with gibberellic acid and kinetin, on the contrary, retarded banana ripening as indicated by higher values for firmness, starch, cellulose and hemicellulose. The results obtained suggest that ripening of banana can be controlled chemically.     

家蝇幼虫营养保健果冻的研制

家蝇Musca domestica L幼虫营养丰富,体内含有多种具有特异生物活性物质,有很好的开发利用价值。以家蝇幼虫为原料,运用正交试验设计对家蝇幼虫营养保健果冻的加工工艺进行研究。结果表明:35%的家蝇幼虫粗提液,15%的白砂糖,0.18%的柠檬酸和1.4%的卡拉胶的工艺参数,可生产具有良好感官和食用品质的家蝇幼虫营养保健果冻;家蝇幼虫粗提液的护色剂使用0.02%抗坏血酸与0.02%柠檬酸,果冻防腐剂使用0.04%山梨酸钾。     

Inactivation of Escherichia coli and Shigella in acidic fruit and vegetable juices by peroxidase systems

AIMS: To study the bactericidal properties of the lactoperoxidase (LPER)-thiocyanate and soybean peroxidase (SBP)-thiocyanate systems at low pH, their efficiency for inactivation of Escherichia coli and Shigella in acidic fruit and vegetable juices, their effect on colour stability of the juices and interaction with ascorbic acid. METHODS AND RESULTS: Three-strain cocktails of E. coli and Shigella spp. in selected juices were supplemented with the LPER or SBP system. Within 24 h at 20 degrees C, the LPER system inactivated both cocktails by > or = 5 log10 units in apple, 2-5 log10 units in orange and < or = 1 log10 unit in tomato juices. In the presence of SBP, browning was significant in apple juice and white grape juice, slight in pink grape juice and absent in orange or tomato juice. Ascorbic acid protected E. coli and Shigella against inactivation by the LPER system, and peroxidase systems significantly reduced the ascorbic acid content of juices. CONCLUSIONS: Our results suggest a different specificity of LPER and SBP for SCN-, phenolic substrates of browning and ascorbic acid in acidic juices. The LPER system appeared a more appropriate candidate than the SBP system for biopreservation of juices. SIGNIFICANCE AND IMPACT OF THE STUDY: This work may open perspectives towards the development of LPER or other peroxidases as biopreservatives in acidic foods.     

Altered production of the active oxygen species is involved in enhanced cytotoxic action of acylated derivatives of ascorbate to tumor cells

Our previous study shows that 6-O-acyl derivatives of L-ascorbic acid inhibits more markedly cell growth of mouse Ehrlich carcinoma than ascorbic acid. The present study shows that 6-O-palmitoyl ascorbic acid but not ascorbic acid prolongs the lifespan of mice into which tumors such as Meth A fibrosarcoma, MM46 mammary carcinoma, Ehrlich carcinoma and sarcoma 180 are implanted. The potentiated cytotoxicity of 6-O-palmitoyl ascorbic acid is not due to an increase in duration time of the cytotoxic action, because 6-O-palmitoyl ascorbic acid is gradually inactivated during contact with tumor cells and exhibits a similar action time curve to that of ascorbic acid as shown by clonal growth assay. Cytotoxicity of 6-O-palmitoyl ascorbic acid is markedly diminished by combined addition of catalase and superoxide dismutase (SOD), as shown by dye exclusion assay, whereas the cytotoxicity was slightly reduced by either enzyme alone but not by the specifically inactivated or heat-denatured enzymes. In contrast, cytotoxicity of ascorbic acid is abolished by catalyse but not SOD. Autooxidation of 6-O-palmitoyl ascorbic acid was not inhibited by catalase plus SOD. The results indicate that cytotoxicity of 6-O-palmitoyl ascorbic acid is attributed at least partly to both hydrogen peroxide (H2O2) and superoxide (O2-.) generated at the early stage. Cytotoxicity of 6-O-palmitoyl ascorbic acid is also appreciably attenuated by singlet oxygen (1O2) scavengers such as hydroquinone, 1,4-diazobicyclo-2,2,2-octane or sodium azide, but not by hydroxyl radical scavengers including butylated hydroxytoluene, D-mannitol, benzoic acid and ethanol. Thus, in contrast to cytotoxicity of ascorbic acid mediated entirely by H2O2 initially generated, acylated ascorbic acid produces a diversity of active oxygen species including H2O2, O2-. and other species secondarily generated via disproportion, which may be additively involved in the enhanced cytotoxic action.     

Effect of exogenous ascorbic acid intake on biosynthesis of ascorbic acid in mice

The effect of exogenous ascorbic acid intake on biosynthesis of ascorbic acid in mice has been studied. After the mice were on diets containing added ascorbic acid for two months, the activities of ascorbic acid synthesizing enzymes in the mouse liver homogenates were measured using L-gulono-gamma-lactone as a substrate. Exogenous ascorbic acid intake (0.5, 1 or 5% in the diet) was able to increase the concentration of ascorbic acid in the blood and to decrease the activities of ascorbic acid synthesizing enzymes in mouse liver. The results suggest that ascorbic acid synthesis was controlled by local regulatory mechanism or by the concentration of ascorbic acid in the hepatic portal blood. Ingestion of dietary erythorbic acid, a stereoisomer of ascorbic acid, had no effect on the activities of ascorbic acid synthesizing enzymes.     

Blood changes in brook trout induced by infection with Aeromonas salmonicida.

Furunculosis was induced in brook trout, Salvelinus fontinalis, by experimental inoculation with Aeromonas salmonicida. Total protein, hemoglobin, sialic acid, fatty acids, triglycerides, cholesterol, inorganic-phosphorus, acid-soluble phosphorus, and lipid-phosphorus decreased in the blood of the infected fish while amino acids, urea, total creatinine, ammonia, and glucose increased. Pyruvic acid, lactic acid, and ascorbic acid values showed no significant change.     

Stopped-flow investigation of the reaction between vitamin E radical and vitamin C in solution

Kinetic study of the reaction between vitamin E radical and vitamin C has been performed. The rates of reaction of vitamin C (ascorbic acid 1, 6-0-stearyl ascorbic acid 2, and 2,6-O-dipalmitoyl ascorbic acid 3) with vitamin E radical (5,7-diisopropyl-tocopheroxyl) in benzene-ethanol (2:1, v/v) solution have been determined spectrophotometrically, using stopped-flow technique. The second-order rate constants obtained are 549 +/- 30 M-1s-1 for 1, 626 +/- 53 M-1s-1 for 2, and 4.84 +/- 1.41 M-1s-1 for 3 at 25.0 degrees C. The result shows that the ascorbic acid ester 2 having a long-alkyl-chain at 6-position is 1.14 times as reactive as the ascorbic acid 1, whereas the ascorbic acid ester 3 substituted at 2-position is only 0.01 times as reactive as the ascorbic acid 1.     

Loss of Ascorbic Acid from Injured Feline Spinal Cord

Feline spinal cord contains 0.97 mM ascorbic acid, as measured by the dinitrophenylhydrazine method. Greater than 90% is maintained in the reduced form. When functioning normally, the CNS conserves its ascorbic acid with a turnover rate of 2% per h. Following contusion injury severe enough to produce paraplegia, ascorbic acid is rapidly lost from injured spinal tissue. Thus, ascorbic acid is decreased 30% by 1 h and 50% by 3 h following injury. Oxidized ascorbic acid is increased at 1, but not 3, h following impact. As a consequence of its many functions in CNS, loss of ascorbic acid may contribute to derangements in spinal cord function following injury.     

Role of ascorbic acid in auxin induced cell elongation

Summary Cytochemical detection of ascorbic acid in cultured root tips of Zea mays shows that dividing cells accumulate ascorbic acid in the cytoplasm. The localization pattern alters in the root tip as the cells begin to elongate. In elongating cells ascorbic acid is distinctly localized on cell walls. Ascorbic acid content per cell inreases with the onset of cell elongation. Fully elongated cells contain fivefold more ascorbic acid than meristematic cells. Cytophotometric analysis reveals a sharp and positive correlation (r=+0.93) between percentage increase in content of ascorbic acid per cell and corresponding increase in cell size at different phases of cell elongation. IAA treatment to the roots raises the content of ascorbic acid per cell with a parallel increase in size of cell. Involvement of ascorbic acid in IAA induced cell elongation is discussed.     

In vitro oxidation of ascorbic acid and its prevention by GSH

The interaction of glutathione (GSH) with ascorbic acid and dehydroascorbic acid was examined in in-vitro experiments in order to examine the role of GSH in protecting against the autoxidation of ascorbic acid and in regenerating ascorbic acid by reaction with dehydroascorbic acid. If a buffered solution (pH 7.4) containing 1.0 mM ascorbic acid was incubated at 37 degrees C, there was a rapid loss of ascorbic acid in the presence of oxygen. When GSH was added to this solution, ascorbic acid did not disappear. Maximum protection against ascorbic acid autoxidation was achieved with as little as 0.1 mM GSH. Cupric ions (0.01 mM) greatly accelerated the rate of autoxidation of ascorbic acid, an effect that was inhibited by 0.1 mM GSH. Other experiments showed that GSH complexes with cupric ions, resulting in in a lowering of the amount of GSH in solution as measured in GSH standard curves. These results suggest that the inhibition of ascorbic acid autoxidation by GSH involves complexation with cupric ions that catalyze the reaction. When ascorbic acid was allowed to autoxidize at 37 degrees C the subsequent addition of GSH (up to 10 mM) did not lead to the regeneration of ascorbic acid. This failure to detect a direct reaction between GSH and the dehydroascorbic acid formed by oxidation of ascorbic acid under this condition was presumably due to the rapid hydrolysis of dehydroascorbic acid. When conditions were chosen, i.e., low temperature, that promote stability of dehydroascorbic acid, the direct reaction between GSH and dehydroascorbic acid to form ascorbic acid was readily detected. The marked instability of dehydroascorbic acid at 37 degrees C raises questions regarding the efficiency of the redox couple between GSH and dehydroascorbic acid in maintaining the concentration of ascorbic acid in mammalian cells exposed to an oxidative challenge.     

The Relationship Between the Phosphorus Status of Subterranean Clover Plants and Losses in Chlorophyll from Detached Leaves: A Basis for the Visual Diagnosis of Phosphorus Deficiency

Leaves detached from subterranean clover plants (Trifolium subterraneumL.), grown in solution cultures at different phosphorus levels,were placed in water and in phosphate solutions. Losses in totalchlorophyll (a+b) of leaves in water were greater the lowerthe previous phosphorus supply to the plants from which theywere detached. In comparable leaves placed in phosphate solutionschlorophyll was maintained at levels at least as high as inleaves from non-deficient plants. For the latter there wereno differences between treated and untreated leaves. The differencesin colour change between treated and untreated leaves, therefore,increased with the deficiency in the phosphorus supply to theplants from which the leaves were detached, thus visibly reflectingtheir phosphorus status. A light intensity of 300 ft c (27 lx) or higher was necessaryfor maximum differences in leaf colour between treated and untreatedleaves, mainly because lower light levels reduced chlorophylllosses from leaves in water. Differences in colour between treated and untreated deficientleaves decreased with their age. However, valid comparisonsbetween treated and untreated leaf tissue could be ensured bythe use of treated and untreated leaflets. It is suggested that the results provide the basis for a simple,direct and visual diagnostic method, requiring no laboratoryequipment. Trifolium subterraneum L., subterranean clover, phosphorus deficiency, chlorophyll     

Ascorbic acid, a familiar small molecule intertwined in the response of plants to ozone, pathogens, and the onset of senescence

Ascorbic acid is a well‐known antioxidant and cellular reductant with an intimate and complex role in the response of plants to ozone. It is clear from a number of studies that sensitivity to ozone is correlated with total ascorbic acid levels, and that a first line of defence against the reactive oxygen species generated in the apoplastic space by ozone is ascorbic acid. For activity, ascorbic acid must be in the fully reduced state. Therefore, both the rate of ascorbic acid synthesis and recycling via dehydroascorbate and monodehydroascorbate reductases are critical in the maintenance of a high ascorbic acid redox state. Active transport of ascorbic acid across the plasma membrane is necessary to achieve reduction of oxidized ascorbic acid by cytoplasm‐localized reductases. It has been known for some time that the chlorotic lesions produced by exposure to ozone are not unlike lesions produced by the hypersensitive response to avirulent pathogen attack. Surprisingly, activation of a defence gene‐signalling network by both ozone and pathogens is influenced by the level of ascorbic acid. Indeed, in addition to acting simply as an antioxidant in the apoplastic space, ascorbic acid appears to be involved in a complex phytohormone‐mediated signalling network that ties together ozone and pathogen responses and influences the onset of senescence.     

In vivo antimutagenic effect of ascorbic acid against mutagenicity of the common antiamebic drug diiodohydroxyquinoline

We have previously shown that the common antiamebic drug diiodohydroxyquinoline (DIHQ) exhibits mutagenic activity in the in vivo micronucleus test in Swiss albino mice. Results of experiments undertaken to study the influence of ascorbic acid (vitamin C) on the mutagenicity of DIHQ in this model system showed that ascorbic acid acts as an antimutagen against DIHQ. The effective antimutagenic doses of ascorbic acid themselves do not show any genotoxic effects in this in vivo system. It will be necessary, however, to elucidate the mechanism of action of ascorbic acid as well as its effects on the therapeutic properties of DIHQ before a practical use of ascorbic acid is contemplated for this purpose.     

Ascorbic acid within chromaffin granules. In situ kinetics of norepinephrine biosynthesis

Ascorbic acid requirements for norepinephrine biosynthesis were investigated in intact bovine chromaffin granules using the physiologic substrate dopamine and a novel coulometric electrochemical detection high pressure liquid chromatography system for ascorbic acid. 10 mM external dopamine, 1 mM Mg-ATP, and 1 mM ascorbic acid produced maximal norepinephrine biosynthesis without granule lysis. When external ascorbic acid was omitted, intragranular ascorbic acid was consumed in a 1:1 ratio with respect to norepinephrine biosynthesis. The initial concentration of intragranular ascorbic acid was 10.5 mM, which was depleted in stepwise fashion to 15 lower concentrations over the range of 9.2-0.2 mM. Chromaffin granules containing these varying concentrations of intragranular ascorbic acid were then incubated with 1 mM exogenous ascorbic acid, and norepinephrine biosynthesis from dopamine was determined. The apparent Km of norepinephrine biosynthesis for intragranular ascorbic acid was 0.57 mM by Eadie-Hofstee analysis and 0.68 mM by Lineweaver-Burk analysis. These data indicate that intragranular ascorbic acid is available and required for norepinephrine biosynthesis, that ascorbic acid is a true co-substrate for dopamine beta-monooxygenase, and that intragranular ascorbic acid is maintained by extragranular ascorbic acid. Continued norepinephrine biosynthesis in granules is dependent on both intragranular and extragranular concentrations of the vitamin. Furthermore, in situ kinetics of dopamine beta-monooxygenase for ascorbic acid may be most accurately determined using intact granules and the true physiologic substrate.     

Ascorbic acid recycling in human erythrocytes is induced by smoking in vivo

Tobacco smoke contains large numbers of radicals that burden the antioxidant defense and, thus, lower plasma antioxidants, in particular vitamin C or ascorbic acid, is commonly observed among smokers. Ascorbic acid recycling describes the process in which ascorbic acid is oxidized to dehydroascorbic acid by various pathways and subsequently reduced back to ascorbic acid intracellularly, e.g., in erythrocytes, thereby preserving the ascorbic acid pool. In humans who are unable to synthesize ascorbic acid, and in smokers in particular, who are prone to oxidation, this process must be very efficient and of great importance. It has previously been reported that isolated erythrocytes subjected to tobacco smoke in vitro had significantly lower ascorbic acid recycling as compared to controls. In contrast to these findings, we now report that freshly isolated erythrocytes from long-term smokers (n = 39) display a significantly increased rate of ascorbic acid recycling in vivo as compared to those isolated from nonsmokers (n = 31; p <.0001). Preliminary data suggests that the increase results from induction of dehydroascorbic acid reductase activity rather than from differences in energy status, glutathione content, or altered transport capacity. The induction of ascorbic acid recycling as a potential adaptation mechanism of the antioxidant defense to oxidative insults is discussed.     

Ascorbate is particularly effective against LDL oxidation in the presence of iron(III) and homocysteine/cystine at acidic pH

Metal-catalyzed LDL oxidation is enhanced by the presence of homocysteine. In this study, the effectiveness of ascorbic acid against low-density lipoprotein (LDL) oxidation by iron(III) and copper(II) in the presence of homocysteine and the main plasma disulfide cystine was investigated. Relative to the degree of LDL oxidation reached in the absence of antioxidants, ascorbic acid was particularly effective against iron-catalyzed LDL oxidation at pH 6.0. This can be explained from its stability under acidic conditions and is likely to be important in ischemia, in inflammation and exhausting exercise. At pH 7.4, an ascorbic acid concentration at least as high as the concentration of homocysteine might be necessary to efficiently inhibit LDL oxidation by iron(III) and copper(II) in the presence of homocysteine and cystine. Histidine increased the efficiency of ascorbic acid as an antioxidant against copper-mediated oxidation in this system. The capacity of homocysteine to regenerate ascorbic acid from dehydroascorbic acid appeared to play a minor role in inhibition of ascorbic acid oxidation by copper as compared to copper chelation by homocysteine.     

Biosynthesis and metabolism of ascorbic acid in plants

The biosynthesis of L‐ascorbic acid in plants differs from that encountered in ascorbic acid‐synthesizing animals. Enzymic details are sparse, but in vivo studies with tracers clearly establish the stereochemical detail of both processes. Examples of each process are found in separate classes of algae. Plants utilize L‐ascorbic acid as the carbon source for the biosynthesis of two important plant acids, oxalic acid and L‐tartaric acid. Here, cleavage of L‐ascorbic acid between carbons 2 and 3 releases the 2 and 4 carbon intermediates. A second L‐tartaric acid‐synthesizing process peculiar to vitaceous plants, i.e., grape, cleaves ascorbic acid between carbons 4 and 5. The physiological significance of these metabolic interconversions is discussed. Other metabolic processes such as the oxidation/reduction properties of L‐ascorbic acid are also considered.