Yield and post-harvest quality of tomato hybrids heterozygous at the loci alcobaça, old gold-crimson or high pigment

The effects of the fruit ripening mutant gene alcoba?a (alc) and color development mutants, old gold-crimson (ogc) and high pigment (hp), on yield and post-harvest quality of tomato fruits were investigated. Five tomato hybrids were obtained by crossing near isogenic lines with Flora-Dade background [Flora-Dade (alc+/alc+ ogc+/ogc+ hp+/hp+), TOM-559 (alc/alc ogc+/ogc+ hp+/hp+), TOM-591 (alc/alc ogc/ogc hp+/hp+), TOM-593 (alc/alc ogc+/ogc+ hp/hp), and TOM-589 (alc/alc ogc/ogc hp/hp)] with the pollen parent line Mospomorist (alc+/alc+ ogc+/ogc+ hp+/hp+). Hybrid fruit was harvested at the breaker stage and stored on shelves at 15oC and 60% relative humidity for 16 days, and then evaluated for firmness, development of red color, and carotenoid contents. The different genotypic combinations at the loci alc, ogc and hp had no effect on fruit yield. The alc+/alc hybrid genotype significantly increased fruit firmness and significantly delayed the development of red color in maturing fruit. Simultaneous usage of ogc+/ogc and hp+/hp promoted an increase in the red color and lycopene content of alc+/alc hybrids, but did not have any additional effect on fruit firmness.     

pH and buffering in the bicinchoninic acid (4,4'-dicarboxy-2,2'-biquinoline) protein assay

The HCO3/CO3(2-) buffer used in the bicinchoninic acid (BCA) protein assay has only weak buffering capacity at the recommended pH (11.25). Consequently the assay is rather sensitive to interference from effectively acid or alkaline samples, particularly in the micro method. Adjustment of pH in these alkaline solutions of high [Na+] is complicated by Na+ errors on the pH electrode. Hence it is recommended to prepare the buffers from known amounts of NaHCO3 and Na2CO3, and to reduce the pH to around 10.7; this offers much better buffering capacity with only a limited reduction in color development.     

Interference of biogenic amines with the measurement of proteins using bicinchoninic acid

The use of bicinchoninic acid (BCA) to measure protein concentrations has received wide acceptance because the reagent is insensitive to many of the buffers, sucrose solutions and detergents used with various tissue and enzyme preparations. However, any compound capable of reducing Cu2+ in an alkaline medium such as biogenic amines will produce a color reaction. The primary objective of this study was to determine whether biogenic amines present in neuronal tissue would interfere with the measurement of protein using the BCA method. Catecholamines were found to produce a linear increase in color of the BCA reagent at concentrations between 1 and 100 nmol/2.1 ml assay volume. Catecholamines appeared to be more sensitive to the BCA reagent than either serotonin or ascorbic acid. Catecholamines at concentrations of 50 nmol/mg of protein or 1 nmol/2.1 ml assay volume or higher will produce significantly (P less than 0.0001) higher color reactions than protein alone. The BCA reagent is not ideal for measuring protein concentrations of intact synaptic vesicles and chromaffin granules since the catecholamine concentrations in these organelles are high enough to increase the color developed by 1.1 to 2.5 times that observed with protein alone. The linearity of the color development produced by catecholamines suggest that BCA could be used to quantitate catecholamine concentrations between 1 and 100 nmol. The BCA reagent will not distinguish between the different catecholamines.     

In vitro evaluation and pregnancy rates after vitrification of in vitro produced bovine embryos

The efficacy of different vitrification solutions to cryopreserve in vitro-produced bovine blastocysts was evaluated based on in vitro development of embryos in culture and on in vivo development of embryos transferred into recipients. In the first experiment, 2 vitrification solutions were compared: propylene glycol + glycerol (Pg + Gly) and ethylene glycol + Ficoll + sucrose (EFS). Differences in the overall development and hatching rates in favor of EFS were found (56.4 vs 33.3% and 35.4 vs 13.3%; P < 0.05). In the second experiment, 3 vitrification solutions were compared: EFS, modified EFS (EFSm) and ethylene glycol + glycerol (Eg + Gly). The vitrification solutions EFSm and Eg + Gly yield higher hatching rates than did EFS (57.7 vs 59.6 vs 35.7%; P < 0.05). The last experiment was designed to compare in vivo 2 vitrification solutions: EFSm and Eg + Gly. There were no differences between them based on the results obtained after transfer (35.2 vs 43.7%). The vitrification solutions EFSm and Eg + Gly have resulted in good pregnancy rates. These results demonstrated that vitrification can be used successfully in the cryopreservation of in-vitro produced bovine embryos, and it might be considered for use in commercial programs.     

The chemical mechanism for Al complexing with delphinidin: A model for the bluing of hydrangea sepals

The blooms of many hydrangea cultivars can be red or blue, with the color depending on the soil pH. This dependence reflects the availability of Al³⁺ to the plant under acidic conditions, as Al³⁺ changes the color of the anthocyanin pigment in hydrangea sepals from red to blue. A chemical model, Al³⁺ and delphinidin in acidic ethanol, was developed to understand the spectral characteristics and bluing of the hydrangea sepals. Delphinidin as its flavylium cation leads to red solutions in the model system. In the presence of Al³⁺, the Al³⁺ removes H⁺ ions from delphinidin, transforming delphinidin's flavylium cation to its blue quinoidal base anion which complexes with the Al³⁺. To further stabilize this complex, a second flavylium cation stacks on top of the complexed quinoidal base anion, creating a bathochromic shift of the cation's spectral signature and accentuating the blue color. This Al³⁺-delphinidin entity forms in adequate concentration for bluing only if there is a sufficient excess of Al³⁺, the exact excess being a function of pH and concentration. The role of Al³⁺ in bluing is not just to form a primary complex with delphinidin, but also to create a template for the stacking of delphinidin (or possibily co-pigments).     

High vanadate interferes with the Fiske-Subbarow determination of inorganic phosphate

We evaluated the possibility that oxyions of vanadium might react with molybdate and, in that manner, interfere with the Fiske-Subbarow colorimetric determination of inorganic phosphate. Phosphate (Pi) standard curves were prepared (0.03-0.30 mumole/ml) in the presence and absence of oxyvanadium solutions (2 X 10(-4) M) prepared from ortho- and metavanadate. Molybdate prepared in 5 N sulfuric acid was added to each standard. Upon addition of a reducing agent to develop color of the phosphomolybdate complex, a less intense color was observed at any given Pi concentration in the presence of oxyvanadium species. The slope of the regression line for the Pi standard curve in the presence of 2 X 10(-4) M oxyvanadium species was markedly depressed. The effect of oxyvanadium was similar when solutions were prepared from ortho- and metavanadate, despite differences in pH of these solutions. In addition, in the final reaction the pH was similar in the presence and absence of oxyvanadium, independent of the source of vanadate used to prepare solutions. Thus, interference by oxyvanadium did not appear to be related to changes in pH of samples containing vanadium oxyions. Interference was concentration dependent and the minimal concentration of vanadium oxyions that interfered was 5 X 10(-5) M. The effects of oxyvanadium (2 X 10(-4) M) on Mg+2-dependent and Na+-K+-ATPase activities in a renal microsomal preparation were then evaluated through the measurement of inorganic phosphate generation. Enzyme activities were determined with and without correction for interference by oxyvanadium with the method of Fiske and Subbarow. A significant artifactual depression of Mg+2 ATPase activity, but not Na+-K+-ATPase activity, was consistently observed when enzyme activities were not corrected for interference by oxyvanadium with the measurement of inorganic phosphate. These data indicate that when effects of high vanadate concentrations (5 X 10(-5) M) on ATP hydrolyzing enzymes are evaluated through changes in Pi generation, artifactual depression of enzyme activity may occur.     

On the specificity of the folin and lowry reagents for amine derivatives.

Reactivities of several amine derivatives with the Folin and Lowry reagents were examined. Tertiary amines reacted with the Folin reagent to produce a blue color, and secondary amines having a 2-hydroxyethyl group reacted with the Folin reagent only in the presence of Cu²⁺, i.e., with the Lowry reagent. On the other hand, primary and quarternary amines and amine N-oxides produced no color with either reagent. Reactivities of tertiary amines were greatly influenced by the nature of the N-substituted groups, and the color yield of those forming stable chelate complexes with metals was strongly inhibited by the presence of Cu²⁺, indicating that the formation of a stable complex with Cu²⁺ reduces the reactivity of tertiary amino nitrogen. The requirement of Cu²⁺ for the color development with secondary amines having a 2-hydroxyethyl group may be due to the formation of weak chelate complex with Cu²⁺.     

Development and validation of an economical uric acid-Fe3+/Fe2+-ferrozine-based colorimetric assay to estimate uric acid level of pure and biological samples

ABSTRACT

This work presents the development and validation of a simple, rapid, and cost-effective spectrophotometric method for quantitative analysis of uric acid in biological samples. The method relies upon uric acid-led reduction of Fe(III) to Fe(II) of sample/standard solutions which stoichiometrically engages ferrozine to form a magenta-colored complex. Different parameters including pH, metal and chelator concentrations, temperature, etc., were optimized for the maximum intensity and stability of the complex. The uric acid concentrations of synthetic/plasma solutions were determined by comparing the color intensity of Fe(ferrozine)₃ ²⁺ complex produced by test solution with the standard curve formed by known uric acid concentrations. The method was validated in accordance with ICH guidelines and subjected to human plasma analysis. The results obtained were compared with a reference (enzymatic) method which revealed that there was no significant difference between the two methods at 95% confidence level. The method is highly specific, precise, linear, accurate, and robust.     

A Consideration of French's Test of Basic Fuchsin for Endo's Medium

In describing a method of testing for the return of color in decolorized fuchsin for use in Endo Medium, French states that variations in hydrogen ion concentration fail to influence the appearance of color in this medium.

Duplications of this test were made using alcoholic and aqueous solutions of fuchsin and both sodium sulfite and sodium bisulfite as decolorizing agents.

In the decolorized alcoholic solutions of fuchsin the color failed to reappear when formalin was added, but a small amount of a weak solution of lactic acid caused the color to return.

Alcoholic solutions of fuchsin failed to decolorize in sodium bisulfite solutions until a few drops of NaOH were added. The color, then, reappeared immediately.

Solutions of peptones to which fuchsin had been added were substituted for the original fuchsin solution. Alcoholic and aqueous solutions of fuchsin were added to equal amounts of a 1% peptone solution. The peptone solutions varied in their hydrogen ion concentration and the results showed that those which were neutral decolorized readily while the more acid solutions were but partially decolorized.

Fuchsin decolorized according to results found in this test, was not satisfactory in the Endo medium, especially in the case of the aqueous solutions of fuchsin.

Experiments which were carried on by other workers and checked with this method all indicated that some acid is necessary to secure the restoration of color.     

Different cellular targets for Cu- and Fe-catalyzed oxidation observed using a Cu-compatible thiobarbituric acid assay

The widely used thiobarbituric acid (TBA) assay for oxidative damage to biomolecules fails in Cu2(+)-containing solutions due to the formation of a cloudy precipitate. The chelation of Cu2+ ions with EDTA or Chelex was investigated. Both prevented precipitate formation, but only Chelex allowed proper color development in the TBA assay. The Chelex modified assay could be adapted to a variety of systems, and was applied to the detection of Cu2+/ascorbate dependent deoxyribose breakdown and oxidative damage in erythrocyte ghosts, lysates and whole cells. Using this method, it was shown that Cu2+/ascorbate caused membrane damage in ghosts but not in whole red blood cells (RBC). Fe3+/ascorbate, on the other hand, caused formation of TBA-reactive products even in whole RBC. When Cu2+ and Fe3+ were presented to isolated hemoglobin as their 1:1 nitrilotriacetate complexes, the protein bound 10-12 cupric ions per molecule, but no ferric ions. It is suggested that oxidative damage catalyzed by copper or iron ions has different cellular targets, determined by the different binding properties of the two metals to various cellular components.     

植物Na+/H+逆向转运蛋白功能及调控的研究进展

Na+/H+逆向转运蛋白是一种调控Na+、H+跨膜转运的膜蛋白,对细胞内Na+的平衡和pH值的调控等活动具有重要作用。该文主要对近年来Na+/H+逆向转运蛋白功能及其调控的研究进展进行概述,着重讨论其在调控离子稳态平衡,液泡pH值大小与花色显现,以及在影响细胞,器官(叶片)发育,盐胁迫信号转导等方面的可能作用。     

Population heterogeneity and color stimulus heterogeneity in agent-based color categorization

Investigating the interactions between universal and culturally specific influences on color categorization across individuals and cultures has proven to be a challenge for human color categorization and naming research. The present article simulates the evolution of color lexicons to evaluate the role of two realistic constraints found in the human phenomenon: (i) heterogeneous observer populations and (ii) heterogeneous color stimuli. Such constraints, idealized and implemented as agent categorization and communication games, produce interesting and unexpected consequences for stable categorization solutions evolved and shared by agent populations. We find that the presence of a small fraction of color deficient agents in a population, or the presence of a “region of increased salience” in the color stimulus space, break rotational symmetry in population categorization solutions, and confine color category boundaries to a subset of available locations. Further, these heterogeneities, each in a different, predictable, way, might lead to a change of category number and size. In addition, the concurrent presence of both types of heterogeneity gives rise to novel constrained solutions which optimize the success rate of categorization and communication games. Implications of these agent-based results for psychological theories of color categorization and the evolution of color naming systems in human societies are discussed.     

Determination of Glucose by a Modification of Somogyi-Nelson Method

Nelson’s arsenomolybdate, the chromogenic reagent in Somogyi–Nelson method, was replaced by Folin–Ciocalteu phenol reagent. The major object was to remove the toxic arsenic compounds from the color reaction system. The color-producing ability of the phenol reagent was considerably lower than that of Nelson’s reagent. However, the modified method was favorably comparable to Somogyi–Nelson method in simplicity, reproducibility and stability of color development. The error in both the modified and Somogyi–Nelson method could be reduced to about one fourth by adding sodium benzoate (final concentration, about 0.5%) to the test solutions.     

Color and toxicity removal following tyrosinase-catalyzed oxidation of phenols

The products of phenol oxidation catalyzed by mushroom tyrosinase (polyphenol oxidase, EC 1.14.18.1) were assessed in terms of their residual color and toxicity. The addition of aluminum sulfate had little effect on the removal of colored products from phenol solutions treated with tyrosinase. Although chitosan was used successfully to remove the color when added before the reaction initiation or after the reaction completion, the required dose of chitosan was lower when it was added after the reaction. In this case, the minimum doses of chitosan required to achieve 90% color removal were proportional to the logarithm of the initial concentration of phenol. The color removal induced by chitosan addition appeared to be the result of chemical interaction followed by a coagulation mechanism. All treated solutions of phenol and chlorophenols, except 2,4-dichlorophenol, had substantially lower toxicities than their corresponding initial toxicities, as measured using the Microtox assay. Chitosan addition significantly enhanced the reduction in toxicity. The toxicities of the phenol solutions treated with tyrosinase were markedly lower than previously reported toxicities of solutions treated with peroxidase enzymes.     

A simplified method for inorganic phosphate determination and its application for phosphate analysis in enzyme assays

A simplified method for inorganic phosphate determination has been developed. The method is sensitive, easy, economic, and applicable for estimation of phosphate released in both enzymatic and nonenzymatic reactions. A mixture of hydrazine sulfate and ascorbic acid was used as the reducing agent and the conditions for the development of the molybdenum blue color were optimized. Thus in the 4.0 ml assay system, 0.4 ml of the reducing agent solution containing 20 mg each of hydrazine sulfate and ascorbic acid per milliliter of 1.0 N H2SO4 gave a rapid optimum color development with absorption maximum at 820 nm. Color development showed a linear relationship up to 10 microg Pi concentration. Thus the method has a 2.5x higher range of Pi estimation than that of the Bartlett method. The molar extinction coefficient at 820 nm was higher than that obtained in the Bartlett procedure. Also the molybdenum blue color formed was stable up to 24 h. Under the standard assay conditions, interference from acid-labile phosphate as in the case of Na+,K+ ATPase was at the minimum. The applicability of the method for assay of microsomal Na+,K+ ATPase and glucose-6-phosphatase was checked in microassays (final volume 0.1 ml) in comparison to the conventional procedures which use 3-4 times higher volumes. Likewise the applicability of the method for phospholipid analysis was compared with that of the conventional Bartlett method. Under both test systems the results obtained by the micromethod were identical to those obtained by the conventional methods. In general the method, which rapidly produces quantitatively molybdenum blue color, not only is rapid economical, and convenient but also has wide applicability.     

Relation of neurological complications of subarachnoid block to some unseen dangers of new techniques

Sterilizing solutions that by accident become mixed into the anesthetic agent are probably the etiological factor producing many of the neurological complications following spinal anesthesia. Use of sterilizing solutions having an intensity of color so great that even one drop in the contents of an ampule will definitely color the anesthetic agent is the criterion of safety.     

RELATION OF NEUROLOGICAL COMPLICATIONS OF SUBARACHNOID BLOCK TO SOME UNSEEN DANGERS OF NEW TECHNIQUES

Sterilizing solutions that by accident become mixed into the anesthetic agent are probably the etiological factor producing many of the neurological complications following spinal anesthesia.Use of sterilizing solutions having an intensity of color so great that even one drop in the contents of an ampule will definitely color the anesthetic agent is the criterion of safety.     

A vacuolar iron transporter in tulip, TgVit1, is responsible for blue coloration in petal cells through iron accumulation

Blue color in flowers is due mainly to anthocyanins, and a considerable part of blue coloration can be attributed to metal-complexed anthocyanins. However, the mechanism of metal ion transport into vacuoles and subsequent flower color development has yet to be fully explored. Previously, we studied the mechanism of blue color development specifically at the bottom of the inner perianth in purple tulip petals of Tulipa gesneriana cv. Murasakizuisho. We found that differences in iron content were associated with the development of blue- and purple-colored cells. Here, we identify a vacuolar iron transporter in T. gesneriana ( TgVit1 ), and characterize the localization and function of this transporter protein in tulip petals. The amino acid sequence of TgVit1 is 85% similar that of the Arabidopsis thaliana vacuolar iron transporter AtVIT1, and also showed similarity to the AtVIT1 homolog in yeast, Ca²⁺-sensitive cross-complementer 1 (CCC1). The gene TgVit1 was expressed exclusively in blue-colored epidermal cells, and protein levels increased with increasing mRNA expression and blue coloration. Transient expression experiments revealed that TgVit1 localizes to the vacuolar membrane, and is responsible for the development of the blue color in purple cells. Expression of TgVit1 in yeast rescued the growth defect of ccc1 mutant cells in the presence of high concentrations of FeSO₄. Our results indicate that TgVit1 plays an essential role in blue coloration as a vacuolar iron transporter in tulip petals. These results suggest a new role for involvement of a vacuolar iron transporter in blue flower color development.     

Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation.

Solutions used for vitrification or rapid cooling of embryos usually contain high concentrations of penetrating cryoprotectants. At these concentrations embryos can tolerate the penetrating cryoprotectants for only short periods of time without damage. This study designed and tested cryoprotectant solutions that combined high polymer concentrations with low penetrating cryoprotectant concentrations. Mouse 2-cell embryo development was not compromised by up to 15-min exposure to 30 wt% solutions of the polymers Ficoll 70,000 MW or dextran 69,000 MW at room temperature. However, our batches of polyvinylpyrrolidone (PVP) 10,000 and PVP 40,000 were embryo-toxic even after extensive dialysis against Milli-Q water. As both Ficoll and dextran contribute to a solution's physical vitrification properties, we formulated vitrifying solutions containing only 11 to 27 wt% ethylene glycol (EG) by including 34 to 49 wt% polymers (27 wt% EG + 34 wt% Ficoll, 27 wt% EG + 34 wt% dextran, 16 wt% EG + 39 wt% Ficoll, or 11 wt% EG + 49 wt% Ficoll, in phosphate-buffered saline (PBS)). Novel solutions were designed for 0.25 ml straw as a viscous matrix for encapsulation of embryos. These yielded high rates of development of 2-cell mouse embryos after rapid cooling and warming (> or = 96% expanded blastocysts in vitro and > or = 62% viable fetuses as assessed on day 15 of gestation in vivo) in all tested solutions. All control 2-cell embryos formed expanded blastocysts in vitro and 78% formed fetuses in vivo. Comparable results were obtained with both 4-cell and 8- to 16-cell mouse embryos. The lower toxicity of Ficoll and dextran may explain why these new solutions gave better results than had previously been reported for solutions containing 7.5% PVP and low concentrations of EG (2 M).     

Vitrification of in vitro produced bovine embryos: in vitro and in vivo evaluations

The efficacy of different vitrification solutions to cryopreserve in vitro produced bovine blastocysts was evaluated based upon in vitro development of embryos in culture and on in vivo development of embryos transferred into recipients. In the first experiment, ethylene glycol + glycerol (Eg + Gly) + different sucrose concentrations were evaluated. There were no significant differences in development rates among solutions. As for hatching, the Eg + Gly + 0.1 M sucrose group had a greater rate as compared with Eg + Gly + 0 M sucrose and Eg + Gly + 0.5 M sucrose groups in the evaluations of Day 6, Day 7 and Day 6 + Day 7 embryos; and, Eg + Gly + 0.3 M sucrose group had a greater rate as compared with the Eg + Gly + 0 M sucrose and Eg + Gly + 0.5 M sucrose groups in evaluations of Day 6 and Day 6 + Day 7 embryos. There were no significant differences in development and hatching rates between Day 6 and 7 in in vitro produced bovine embryos within each treatment group. There were significant differences in nuclei number after vitrification between Eg + Gly + 0.1 M and Eg + Gly + 0 M sucrose groups and the Eg + Gly + 0.5 M sucrose group. Pregnancy after 60 days of transfer and calving rates showed a difference between in vivo produced embryos freshly transferred and in vitro produced embryos vitrified with Eg + Gly + 0.3 M. There were no significant differences in gestation length and sex ratio between treatments. As for birth weight, there were significant differences between fresh in vivo produced embryos and all treatments of in vitro produced embryos. There were significant differences in dystocial parturition between in vivo produced embryos and all treatments with in vitro produced embryos. These results demonstrate that vitrification can be used successfully in the cryopreservation of in vitro produced bovine embryos, and that it might be considered for use in commercial programs.