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Lactic acid bacteria in the quality improvement and depreciation of wine.

The winemaking process includes two main steps: lactic acid bacteria are responsible for the malolactic fermentation which follows the alcoholic fermentation by yeasts. Both types of microorganisms are present on grapes and on cellar equipment. Yeasts are better adapted to growth in grape must than lactic acid bacteria, so the alcoholic fermentation starts quickly. In must, up to ten lactic acid bacteria species can be identified. They belong to the Lactobacillus, Pediococcus, Leuconostoc and Oenococcus genera. Throughout alcoholic fermentation, a natural selection occurs and finally the dominant species is O. oeni, due to interactions between yeasts and bacteria and between bacteria themselves. After bacterial growth, when the population is over 10(6) CFU/ml, malolactic transformation is the obvious change in wine composition. However, many other substrates can be metabolized. Some like remaining sugars and citric acid are always assimilated by lactic acid bacteria, thus providing them with energy and carbon. Other substrates such as some amino acids may be used following pathways restricted to strains carrying the adequate enzymes. Some strains can also produce exopolysaccharides. All these transformations greatly influence the sensory and hygienic quality of wine. Malic acid transformation is encouraged because it induces deacidification. Diacetyl produced from citric acid is also helpful to some extent. Sensory analyses show that many other reactions change the aromas and make malolactic fermentation beneficial, but they are as yet unknown. On the contrary, an excess of acetic acid, the synthesis of glucane, biogenic amines and precursors of ethylcarbamate are undesirable. Fortunately, lactic acid bacteria normally multiply in dry wines; moreover some of these activities are not widespread. Moreover, the most striking trait of wine lactic acid bacteria is their capacity to adapt to a hostile environment. The mechanisms for this are not yet completely elucidated. Molecular biology has provided some explanations for the behaviour and the metabolism of bacteria in wine. New tools are now available to detect the presence of desirable and undesirable strains. Even if much remains unknown, winemakers and oenologists can nowadays better control the process. By acting upon the diverse microflora and grape musts, they are more able to produce healthy and pleasant wines.

Carbohydrate Sequence↗

Capillary zone electrophoresis as a tool to monitor the last stages of the degradation of water-sensitive polymers.

In order to monitor the formation of the water-soluble by-products from chain-scission of degradable polymers used in the biomedical field, four capillary electrophoresis methods are discussed with the aim of giving the limits and performance for each. Three of them (electroosmotic flow reversal by dynamic adsorption of a polycation, multilayer polyelectrolyte coatings and physical binding of polyethylene oxide) are based on the use of dynamic coatings onto the inner surface of a fused silica capillary, a simple means to adapt performance to specific separations via modification and control of the electroosmotic flow of fused capillary. Using oligomers of lactic acid considered as standards the methods are compared. Other examples of ester-containing macromolecules (poly(hydroxybutyrate)), as well as degradable polyanions are described, namely N-acetylneuraminate polymer and poly(beta-malic acid).

Biodegradation, Environmental↗

Terminal differentiation in the avian uropygial gland. Accumulation of fatty acid synthase and malic enzyme in non-dividing cells.

The secretory tissue of the uropygial gland is of the holocrine type, containing both dividing progenitor cells and lipid-filled differentiated cells. In this study, we examined the relationship between cell division and differentiation. The location of dividing cells was determined by autoradiography of tissue sections from ducklings injected intra-abdominally with 3H-thymidine. Only cells on the basal lamina of the tubules contained labeled nuclei. Dividing cells were distributed uniformly over the length of the tubules. Over the next five days, most of the labeled cells migrated to the lumen of the tubules and disappeared. Cells containing the "lipogenic" enzymes, fatty acid synthase and malic enzyme, were localized either immunocytochemically using affinity-purified antibodies or cytochemically using a specific assay for malic enzyme activity. Fatty acid synthase and malic enzyme were undetectable in dividing basal cells but present at high levels in differentiating and differentiated cells. Thus, basal cells lying along the basal lamina of the tubules were replacing lipid-laden cells that were continually sloughed into the lumens of the tubules. The signals for differentiation and enzyme accumulation appear to be linked to one another and to cessation of cell division.

Aging↗

CRASSULACEAN ACID METABOLISM: Molecular Genetics.

Crassulacean acid metabolism (CAM) is an adaptation of photosynthesis to limited availability of water or CO2. CAM is characterized by nocturnal CO2 fixation via the cytosolic enzyme PEP carboxylase (PEPC), formation of PEP by glycolysis, malic acid accumulation in the vacuole, daytime decarboxylation of malate and CO2 re-assimilation via ribulose-1,5-bisphosphate carboxylase (RUBISCO), and regeneration of storage carbohydrates from pyruvate and/or PEP by gluconeogenesis. Within this basic framework, the pathway exhibits an extraordinary range of metabolic plasticity governed by environmental, developmental, tissue-specific, hormonal, and circadian cues. Characterization of genes encoding key CAM enzymes has shown that a combination of transcriptional, posttranscriptional, translational, and posttranslational regulatory events govern the expression of the pathway. Recently, this information has improved our ability to dissect the regulatory and signaling events that mediate the expression and operation of the pathway. Molecular analysis and sequence information have also provided new ways of assessing the evolutionary origins of CAM. Genetic and physiological analysis of transgenic plants currently under development will improve our further understanding of the molecular genetics of CAM.

Journal Article↗

[Structuro-kinetic organization of the tricarboxylic acid cycle in the active functioning of mitochondria].

Taking into account structural and functional organization of mitochondrial processes it has been shown that at active work there functions in mitochondria an accelerated mechanism of succinic acid formation via coupling of glutamate-oxalacetate transaminase and alpha-ketoglutaratdehydrogenase. This way is closed up into a cycle with the participation of cytosol transaminases which support influx of glutamate, pyruvate and malic acid into mitochondria. When provision of the mitochondria with the substrate proceeds along the transaminase pathway the initial slow region of the tricarboxylic acid cycle is omitted. Thus at active work a faster course is selected. It permits realization of the advantages of succinate dehydrogenase high activity and of oxidation efficiency of succinic acid generated in mitochondria which is essentially higher than that under oxidation of succinic acid and even more of other substrates of the tricarboxylic acid cycle.

Citric Acid Cycle↗

Evidence for a Functional myo-Inositol Oxidation Pathway in Lilium longiflorum Pollen.

Addition of myo-inositol to pentaerythritol-based germination media repressed the conversion of d-[1-(14)C]glucose to labeled uronosyl and pentosyl units of tube wall pectic substance in lily pollen (Lilium longiflorum Thunb.). Conversion of d-[1-(14)C]glucose to labeled glucosyl, galactosyl, and rhamnosyl units was unaffected. The reverse experiment, addition of d-glucose to pentaerythritol-based media, failed to affect the conversion of myo-[2-(3)H]inositol to uronosyl and pentosyl units although the flow of label into products of myo-inositol-linked glucogenesis was blocked. Results of these experiments are discussed in terms of a functional myo-inositol oxidation pathway.d-[1-(14)C]Glucose-labeled pollen tubes contain a labeled, 70% ethyl alcohol-soluble, acidic compound whose formation is blocked by the myo-inositol antagonist, 2-O,C-methylene-myo-inositol (Chen et al. 1977 Plant Physiol. 59: 658). This compound has been identified as l-malic acid.

Journal Article↗

Production of gentamicins by Micromonospora purpurea.

The natural medium contained the following ingredients (g/l): glucose 8.0, or black strap molasses (treated with 0.2--0.3 g/l EDTA) 12.0, fodder yeast (50.0% total nitrogen) 2.0, or folder yeast (40.0% total nitrogen) 6.0, or yeast extract 8.0, or tryptone 8.0, and CaCO3 1.0. Treated black strap molasses with EDTA and fodder yeast proved to be effective in the fermentative production of gentamicins. The most suitable chelating agent was EDTA in the form of disodium for the treatment of Komombo molasses in a concentration of 0.2--0.3 g/l, while potassium ferrocyanide and methylene blue had depressing effects on the production of gentamicins. The most effective carbon source, present in Egyptian black strap molasses, was glucose. Addition of glucose to the medium was preferable at the beginning of the fermentation process. Trace elements present in molasses were very essential for the microbial growth and biosynthesis of gentamicins as proved when molasses ash was added to the natural medium. Organic nitrogen sources were more suitable than inorganic nitrogen sources for the production of gentamicins by Micromonospora purpurea. The microorganism utilized the synthetic medium, but the antibiotic yields were less than those produced in the natural medium. The synthetic medium exhibited stimulatory effects of certain amino acids, organic acids, vitamins, and purine and pyrimidine bases on the fermentative production of gentamicins. Therefore, the ingredients increasing yields of gentamicins were mainly phenylalanine, iso-leucine, lysine, methionine, leucine, arginine, glycine, beta-alanine, cystine, tryptophan, malic acid, maleic acid, cobalamin, folic acid, riboflavin, vitamin B1, vitamin B6, biotin, nicotinamide, uracil, adenine, guanine, and adenosine. Trace elements (Co, Mo, Fe, Cu, Zn, and Mn) exhibited their important role on the biosynthesis and production of gentamicins by Micromonospora purpurea.

Carbon↗

Efficacy of selected acidulants in pureed green beans inoculated with pathogens (Escherichia coli O157:H7 and Listeria monocytogenes).

Studies were conducted to evaluate the combined effect of selected acidulants (acetic, citric, malic, and phosphoric acid) and heat on foodborne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes) in pureed green beans. To establish a consistent reference point for comparison, the molar concentrations of the acids remained constant while the acid-to-puree ratio, titratable acidity, and undissociated acid were either measured or calculated for a target acidified green beans at a pH of 3.8, 4.2, and 4.6. The D-values at 149 degrees F were used as the criteria for acid efficacy. Generally, acetic acid (puree, pH 3.8 and 4.2) represented the most effective acid with comparatively low D-values irrespective of the target microorganism. A 10-s heating at 149 degrees F inactivated approximately 10(6) CFU/ml of E. coli O157:H7 in pureed beans at pH 3.8. The efficacy of acetic acid is likely related to the elevated percent titratable acidity, undissociated acid, and acid-to-puree ratio. The effectiveness (which in this study represents the combined effect of acid and heat) of the remaining acids (citric, malic, and phosphoric) at puree pH values of 3.8 and 4.2 were statistically insignificant (alpha = 0.05). Surprisingly, acetic acid (puree, pH 4.6) appeared to be the least effective as compared to the other acids tested (citric, malic, and phosphoric) especially on E. coli O157:H7 cells, while L. monocytogenes had a similar resistance to all acids at puree pH 4.6. With the exception of citric acid (pH 3.8), acetic acid (pH 4.6), and malic acid (pH 3.8 and 4.6), which were statistically insignificant (P > 0.05), the D-values for L. monocytogenes were statistically different (P < or = 0.05) and higher than the D-values for E. coli under similar experimental conditions. A conservative process recommendation (referred to as the "safe harbor" process) was found sufficient and applicable to pureed green beans for the pH range studied.

Acids↗

CO(2)-concentrating: consequences in crassulacean acid metabolism.

The consequences of CO(2)-concentrating in leaf air-spaces of CAM plants during daytime organic acid decarboxylation in Phase III of CAM (crassulacean acid metabolism) are explored. There are mechanistic consequences of internal CO(2) partial pressures, p(i)(CO(2)). These are (i) effects on stomata, i.e. high p(i)(CO(2)) eliciting stomatal closure in Phase III, (ii) regulation of malic acid remobilization from the vacuole, malate decarboxylation and refixation of CO(2) via Rubisco (ribulose bisphosphate carboxylase/oxygenase), and (iii) internal signalling functions during the transitions between Phases II and III and III and IV, respectively, in the natural day/night cycle and in synchronizing the circadian clocks of individual leaf cells or leaf patches in the free-running endogenous rhythmicity of CAM. There are ecophysiological consequences. Obvious beneficial ecophysiological consequences are (i) CO(2)-acquisition, (ii) increased water-use- efficiency, (iii) suppressed photorespiration, and (iv) reduced oxidative stress by over-energization of the photosynthetic apparatus. However, the general potency of these beneficial effects may be questioned. There are also adverse ecophysiological consequences. These are (i) energetics, (ii) pH effects and (iii) Phase III oxidative stress. A major consequence of CO(2)-concentrating in Phase III is O(2)-concentrating, increased p(i)(CO(2)) is accompanied by increased p(i)(O(2)). Do reversible shifts of C(3)/CAM-intermediate plants between the C(3)-CAM-C(3) modes of photosynthesis indicate that C(3)-photosynthesis provides better protection from irradiance stress? There are many open questions and CAM remains a curiosity.

Carbon Dioxide↗

Biochemical changes during the preservation stage of ripe olive processing.

The influences of initial sodium chloride (6% and 0% w/v in tap water) and acetic acid concentrations (0.3%, and 0.6% v/v), use of starter culture, and aerobic versus anaerobic conditions on the biochemical changes that take place throughout the preservation stage of ripe olive processing were investigated. Glucose, fructose and sucrose were completely consumed during preservation. Mannitol and malic acid were metabolized only in the presence of lactic acid bacteria or oxidative yeast (aerobic treatment). The main metabolites produced were lactic and acetic acid in aerobic or anaerobic treatments inoculated with Lactobacillus plantarum. Methanol and ethanol were present in all the brines although in a lower concentration when conditions were aerobic. Thus, induced lactic fermentation led to the most efficient utilization of carbohydrates and yielded the most suitable physicochemical characteristics for ripe olive preservation.

Chemical Phenomena↗

Bulk and surface modifications of polylactide.

This article reviews various methods of modifying the bulk and surface properties of poly(lactic acid) (PLA) so that the polymer may be used as a drug carrier in a drug delivery system (DDS) and as a cell scaffold in tissue engineering. Copolymerization of lactide with other lactone-type monomers or monomers with functional groups such as malic acid, copolymerization of lactide with macromolecular monomer such as poly(ethylene glycol) (PEG) or dextran, as well as blending polylactide and natural derivatives and other methods of bulk modification are discussed. Surface modifications of PLA-type copolymers, such as surface coating, chemical modification, and plasma treatment are described. Cell culture technology proves the efficiency of bulk and surface modification and the potential application of PLA in tissue engineering.

Absorbable Implants↗

An improved reversed-phase liquid chromatographic method for the analysis of low-molecular mass organic acids in plant root exudates.

Reversed-phase column liquid chromatography (RPLC) was used for the separation and quantification of 10 low-molecular-mass organic acids (malic, malonic, lactic, acetic, maleic, citric, cis-aconitic, succinic, fumaric, and trans-aconitic) in plant root exudates. A mobile phase of 93% 25 mM KH2PO4 at pH 2.5 and 7% methanol at a flow-rate of 1 ml min(-1) resolved all 10 acids in 15 min on a C18 column. Experiments demonstrated a significant (P < 0.05) effect of sample pH on detector response, with peak heights being significantly lower at pH 6.0 compared with pH 2.5, but peak area showed no significant difference. At pH 8.0 and above, both peak height and area differed significantly from injections made at pH 2.5. Limits of detection (LOD) for the 10 acids ranged from 0.05 to 24 microM. Finally, the improved method was applied for the analysis of root exudates from soil cultured field pea, Banksia attenuata, white lupin, and chickpea.

Acids↗

Multigram synthesis of the C29-C51 subunit and completion of the total synthesis of altohyrtin C (spongistatin 2).

A multigram synthesis of the C29-C51 subunit of altohyrtin C (spongistatin 2) has been accomplished. Union of this intermediate with the C1-C28 fragment and further elaboration furnished the natural product. Completion of the C29-C51 subunit began with the aldol coupling of the boron enolate derived from methyl ketone 8 and aldehyde 9. Acid-catalyzed deprotection/cyclization of the resulting diastereomeric mixture of addition products was conducted in a single operation to afford the E-ring of altohyrtin C. The diastereomer obtained through cyclization of the unwanted aldol product was subjected to an oxidation/reduction sequence to rectify the C35 stereocenter. The C45-C48 segment of the eventual triene side chain was introduced by addition of a functionalized Grignard reagent derived from (R)-glycidol to a C44 aldehyde. Palladium-mediated deoxygenation of the resulting allylic alcohol was followed by adjustment of protecting groups to provide reactivity suitable for the later stages of the synthesis. The diene functionality comprising the remainder of the C44-C51 side chain was constructed by addition of an allylzinc reagent to the unmasked C48 aldehyde and subsequent dehydration of the resulting alcohol. Completion of the synthesis of the C29-C51 subunit was achieved through conversion of the protected C29 alcohol into a primary iodide. The synthesis of the C29-C51 iodide required 44 steps with a longest linear sequence of 33 steps. From commercially available tri-O-acetyl-d-glucal, the overall yield was 6.8%, and 2 g of the iodide was prepared. The C29-C51 primary iodide was amenable to phosphonium salt formation, and the ensuing Wittig coupling with a C1-C28 intermediate provided a fully functionalized, protected seco-acid. Selective deprotection of the required silicon groups afforded an intermediate appropriate for macrolactonization, and, finally, global deprotection furnished altohyrtin C (spongistatin 2). This synthetic approach required 113 steps with a longest linear sequence of 37 steps starting from either tri-O-acetyl-d-glucal or (S)-malic acid.

Aldehydes↗

Histochemical Compartmentation of Photosynthesis in the Crassulacean Acid Metabolism Plant Crassula falcata.

The succulent leaf of the obligate Crassulacean acid metabolism plant Crassula falcata comprises two distinct types of parenchyma. The peripheral tissue is dark green, whereas the central tissue is relatively colorless. We have investigated whether the conventional interpretation of Crassulacean acid metabolism-simply, temporal separation of light and dark CO(2) fixation within individual cells-is sufficient. Ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) and chlorophyll, indicating the photosynthetic-carbon-reduction pathway, were concentrated in peripheral tissue. Specific activities of P-enolpyruvate carboxylase (4.1.1.31) and of NAD(+)-malic enzyme (1.1.1.39), indicating capacity for dark CO(2) fixation and release, respectively, were high in both types of parenchyma. Measured directly as malic acid decline at the beginning of the photoperiod, CO(2) "storage" occurred in both tissues. These data indicate that there is a spatial component to Crassulacean acid metabolism in C. falcata.

Journal Article↗

Quantitative determination of perfluorooctanoic acid ammonium salt in human serum by high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry.

A sensitive, specific, accurate and reproducible analytical method was developed and validated to quantify perfluorooctanoic acid (PFOA) in human serum. After initial extraction with an ion-paring reagent, the procedure for quantifying PFOA is based on high-performance liquid chromatography (HPLC) interfaced to negative ion tandem mass spectrometry, operating in selected ion monitoring mode. The retention times of PFOA and its internal standard (D,L-malic acid) were 5.85 and 1.70 min, respectively. The assay was linear over the range 0-500 ng/mL, with a lower limit of quantification (LOQ) of 25 ng/mL, and with a coefficient of variation (CV) of 7.3%. The lower limit of detection (LOD) was assessed as 10 ng/mL. The overall precision and accuracy were assessed on three different days. The within- and between-day precision was < or =9.7 and 6.8%, respectively, and the accuracy was in the range 96-114%. The mean extracted recovery assessed at three different concentrations (100, 250, and 500 ng/mL) was always more than 85%. With this method no derivatization procedure was needed, thus avoiding possible thermal and chemical decomposition reactions of PFOA. The assay was applied to quantify perfluorooctanoic acid in serum from employees exposed to fluorochemicals commonly used in industrial applications for polymer production. The quantitative results for PFOA blood levels were found to vary between 100 and 982 ng/mL.

Caprylates↗

Biochemical response of squirrel monkeys to ozone.

Biochemical studies were performed on blood and lung tissue of squirrel monkeys (Saimiri sciureus) following acute exposure to 0.75 ppm ozone (O3) for 4 h/d for 4 consecutive days. One group of animals was sacrificed at the end of the last exposure day and another group was sacrificed 4 d later after the last exposure. Evidence was sought for oxidation-induced changes known to occur in rodents when high levels of O3 are inhaled. A significant increase in red blood cell membrane fragility was observed, as well as significant decreases in red blood cell glutathione and erythrocyte acetylcholinesterase; however, the red blood cell enzymes, lactic acid dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G6PDH) were not changed significantly. Lung tissue analysis showed that lipid peroxidation was markedly increased and tissue vitamin E levels were significantly decreased. The tissue enzymes G6PDH, glutathione reductase, and LDH significantly increased in activity. No significant changes were seen in either superoxide dismutase or malic acid dehydrogenase. The results of this experiment indicate that O3, or reaction products resulting from O3-tissue interaction in the lung, pass the air-blood barrier and are capable of producing biochemical changes in blood as well as in lung tissue.

Acetylcholinesterase↗

Determination of carboxylic acids, sugars, glycerol and ethanol in wine and grape must by ion-exchange high-performance liquid chromatography with refractive index detection.

Method to determine the major carboxylic acids, sugars, glycerol and ethanol in wine and grape must was developed using an ion-exchange column and refractive index detector. A solid-phase extraction method with a strong anion exchanger was used to determine these compounds in sweet wines and in grape musts. With this method it is possible to determine malic acid in sweet wines and in grape musts without interference from sugars. This high-performance liquid chromatographic method was compared with standard methods of analysis. There was good agreement in the accuracy and precision of the compared methods.

Carbohydrates↗