Physiological and biochemical response of harvested plum fruit to oxalic acid during ripening or shelf-life

The effect of oxalic acid application on plum fruit (Prunus salicina cv. ‘Damili’) ripening properties during storage or shelf-life was determined. The fruits were dipped for 3 min in solutions containing 5 mmol/L oxalic acid and then were packed into polyethylene bags and stored at 25 °C for 12 days, or at 2 °C for 20 days and subsequently at 25 °C for 12 days. Ethylene production, fruit firmness, contents of pectin and anthocyanin, specific activities of polygalacturonase (PG), pectin methylesterase (PME) and phenylalanine ammonia lyase (PAL), and chlorophyll fluorescence (Fv/Fm) were measured. The application of oxalic acid reduced ethylene production and delayed softening of plum fruit. The inhibition of softening was associated with decreased PG and PME activities; that is, the retardation of pectin solubilization/degradation. During storage or shelf-life, flesh reddening and anthocyanin synthesis were significantly inhibited in oxalic acid-treated plum fruit, accompanied with decreased PAL activity. Furthermore, it was found that variable:maximalchlorophyll fluorescence (Fv/Fm), an indicator of ripening, senescence or stress injury of fruit and vegetable, decreased much more slowly in oxalic-treated plum fruits than in control fruits. Thus, oxalic acid treatment can be an effective means to extend the shelf life of plum fruit.     

Identification and quantification of phenols, carotenoids, and vitamin C from papaya (Carica papaya L., cv. Maradol) fruit determined by HPLC-DAD-MS/MS-ESI

Recent studies have demonstrated that vitamin C, phenols, and carotenoids are bioactive compounds that protect the body from oxidative stress, reducing the risk of cardiovascular diseases and some types of cancer. Qualitative and quantitative analysis of the major phytochemicals found in papaya fruit flesh and skin (Carica papaya L., cv Maradol) was conducted in four stages of ripeness, using high-performance liquid chromatography mass spectrometry. Phenolic compounds identified in the fruit skin tended to decrease with ripening. The compounds identified were ferulic acid (277.49 to 186.63 mg/100 gDW), p-coumaric acid (229.59 to 135.64 mg/100 gDW), and caffeic acid (175.51 to 112.89 mg/100 gDW). The following carotenoids, along with vitamin C, increased in flesh with ripening; lycopene (0.36 to 3.40 mg/100 gDW), ??-criptoxanthin (0.28 to 1.06 mg/100 gDW), ??-carotene (0.23 to 0.50 mg/100 gDW), and vitamin C (25.07 to 58.59 mg/100 gDW). These results indicate that stage of ripeness significantly influences the contents of bioactive compounds in papaya fruit.     

Effect of oxalic acid on control of postharvest browning of litchi fruit

Litchi (Litchi chinensis Sonn.) fruit, cv. Huaizhi, was treated with 2 and 4 mM oxalic acid and stored at room temperature to investigate the effect of oxalic acid on pericarp browning. The results showed that the pericarp browning indices of the fruit, treated with both oxalic acid concentrations, were significantly lower than that of the control, due to increase of membrane integrity, inhibition of anthocyanin degradation, decline of oxidation, and maintanance of relatively low peroxidase activity in the fruit during storage. It appears that application of oxalic acid can effectively control the pericarp browning of litchi fruit during postharvest storage.     

Limiting the deterioration of mango fruit during storage at room temperature by oxalate treatment

Effects of oxalate on the incidence of decay and ripening in mango fruit, and its physiological effects on the peel and flesh of mango were investigated after mango fruit (Mangifera indica L.) were dipped in different oxalate solutions for 10 min and then stored at 25 °C. Oxalate application decreased the incidence of decay and delayed the ripening process in mango fruit during storage. Potassium oxalate treatment resulted in increased activities of peroxidase (POD) in both the peel and the flesh and polyphenol oxidase (PPO) in the peel, without activation of phenylalanine ammonia-lyase activity, and elevated total phenolic content in the peel. The physiological effects of oxalate in increasing activities of POD and PPO and elevating total phenolic level could be involved in induced resistance of mango fruit against postharvest disease. Oxalate application could be a promising method to suppress postharvest deterioration and extend the useful shelf-life of mangoes.     

Increased milk levels of transforming growth factor-alpha, beta1, and beta2 during Escherichia coli-induced mastitis

Among the gram-negative bacteria that cause mastitis, Escherichia coli are the most prevalent. The innate immune system provides initial protection against E. coli infection by detecting the presence of the foreign pathogens and by mounting an inflammatory response, the latter of which is mediated by cytokines such as IL-1β, IL-8, and tumor necrosis factor (TNF)-α. Although changes in these cytokines during mastitis have been well-described, it is believed that other mediators moderate mammary gland inflammatory responses as well. The growth factors/cytokines transforming growth factor (TGF)-α, TGF-β1, and TGF-β2 are all expressed in the mammary gland and have been implicated in regulating mammary gland development. In other tissues, these growth factors/cytokines have been shown to moderate inflammation. The objective of the current study was to determine whether TGF-α, TGF-β1, and TGF-β2 milk concentrations were altered during the course of E. coli-induced mastitis. The contralateral quarters of 11 midlactating Holstein cows were challenged with either saline or 72 cfu of E. coli, and milk samples were collected. Basal milk levels of TGF-α, TGF-β1, and TGF-β2 were 98.81 ± 22.69 pg/mL, 3.35 ± 0.49 ng/mL, and 22.36 ± 3.78 ng/mL, respectively. Analysis of whey samples derived from E. coli-infected quarters revealed an increase in milk levels of TGF-α within 16 h of challenge, and these increases persisted for an additional 56 h. Elevated TGF-β1 and TGF-β2 milk concentrations were detected in E. coli-infected quarters 32 h after challenge, and these elevations were sustained throughout the study. Because TGF-α, TGF-β1, and TGF-β2 have been implicated in mediating inflammatory processes, their induction during mastitis is consistent with a role for these molecules in mediating mammary gland host innate immune responses to infection.     

Identification and quantification of major phenolic compounds from mango (Mangifera indica, cv. Ataulfo) fruit by HPLC–DAD–MS/MS-ESI and their individual contribution to the antioxidant activity during ripening

Mango (Mangifera indica L.) is an economically important fruit throughout the world. ‘Ataulfo’ mango, a leading cultivar in Mexico, has the highest content of phenolic compounds among several commercial varieties of mango. However, the individual identification and antioxidant contribution of these phenols during ripening of mango fruit is unknown. Qualitative and quantitative analysis of the major phenolic compounds found in ‘Ataulfo’ mango fruit pulp was conducted in four stages of ripeness, using high-performance liquid chromatography coupled to mass spectrometry. The antioxidant contribution of each of the major phenolic compounds was calculated. The major compounds identified were chlorogenic acid (28–301 mg/100 g DW), gallic acid (94.6–98.7 mg/100 g DW), vanillic acid (16.9–24.4 mg/100 g DW), and protocatechuic acid (0.48–1.1 mg/100 g DW). The antioxidant contribution of the four phenolic acids increased during ripening. Gallic acid accounted for the highest contribution (39% maximum value), followed by chlorogenic acid (21% maximum value). This could indicate that these phenolic compounds may have an important role in the antioxidant metabolism in ‘Ataulfo’ mango fruit during ripening, and promoting health benefits to consumers.     

Effect of pre- and postharvest salicylic acid treatment on ethylene production,fungal decay and overall quality of Selva strawberry fruit

The effect of salicylic acid (SA) treatment at different concentrations and growth stages of strawberry (Fragaria ananassa cv. Selva) fruit on postharvest ethylene production, fungal decay and overall quality index was studied. SA at all concentrations effectively reduced fruit ethylene production and fungal decay and retained overall quality. Treatment of plants at vegetative stage and fruit development stage followed by postharvest treatment of fruits with 1 and 2 mmol L⁻¹ was the most effective strategy, whilst with decrease in treatment time the effects of treatment decreased. Single stage treatment strategy of fruits with 2 mmol L⁻¹ SA at postharvest stage was most effective. Postharvest treatment with 4 mmol L⁻¹ SA slightly damaged the fruits and was less effective than 2 mmol L⁻¹ in retaining fruit quality.     

Antioxidant,anticholinesterase and antimicrobial constituents from the essential oil and ethanol extract of Salvia potentillifolia

The essential oil of Salvia potentillifolia was analysed by GC and GC–MS. Totally, 123 components were detected in both hydrodistilled and steam-distilled oils, α- and β-pinenes being major compounds. The antioxidant activities were determined by using complementary tests, namely, DPPH radical-scavenging, β-carotene-linoleic acid and reducing power assays. The ethanol extract also showed better activity (IC₅₀ = 69.4 ± 0.99 μg/ml) than that of BHT in the DPPH system, and showed great lipid peroxidation inhibition in the β-carotene-linoleic acid system (IC₅₀ = 30.4 ± 0.50 μg/ml). The essential oil showed meaningful butyrylcholinesterase activity (65.7 ± 0.21%), and α-pinene showed high acetylcholinesterase inhibitory activity (IC₅₀ = 86.2 ± 0.96 μM) while β-pinene was inactive. Antimicrobial activity was also investigated on several microorganisms, and the essential oil showed high activity against Bacillus subtilis and B. cereus. It also exhibited remarkable anticandidal activity against Candida albicans and C. tropicalis with MIC values of 18.5 and 15.5 μg/ml, respectively, while α- and β-pinenes showed moderate activity.     

Flavonol glycosides from whole cottonseed by-product

Cottonseeds are fed to high-producing dairy cows as a source of fat and highly-digestible fibre. Seven flavonol glycosides have been identified from whole cottonseed by-product. Their structures were established as quercetin 3-O-{β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside} (1), kaempferol 3-O-{β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside} (2), quercetin 3-O-[β-d-apiofuranosyl-(1 → 2)-β-d-glucopyranoside] (3), quercetin 3-O-β-d-glucopyranoside (4), kaempferol 3-O-[α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside] (5), quercetin 3-O-[α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside] (6), and kaempferol 3-O-α-l-rhamnopyranoside (7). Gallic acid (8) and 3,4-dihydroxybenzoic acid (9) were also found. All structures were elucidated by ESI-MS and NMR spectroscopic methods. Total polyphenols were assayed by the Folin–Ciocalteu method.     

氯吡苯脲浸泡处理延缓采后芒果成熟和软化的生理机制

以采后‘贵妃’芒果为试材,使用10 mg/L氯吡苯脲（forchlorfenuron,CPPU）溶液对果实浸泡处理10 min,晾干后在室温（25 ℃）条件下贮藏8 d,研究CPPU浸泡处理对芒果果实贮藏期间成熟与软化相关生理指标的影响。结果显示：CPPU浸泡处理明显抑制了芒果果实的呼吸作用并推迟了果肉色泽、硬度、可溶性固形物质量分数和可滴定酸质量分数的变化;CPPU浸泡处理有效抑制了果实1-氨基环丙烷-1-羧酸合成酶和1-氨基环丙烷-1-羧酸氧化酶活性,导致乙烯释放量降低;此外,CPPU浸泡处理显著抑制了果实多聚半乳糖醛酸酶、果胶甲酯酶、β-半乳糖苷酶等果胶水解酶活性变化,并较大程度抑制了果胶多糖的增溶与解聚作用,故而延缓了果实的软化进程。结论：CPPU浸泡处理能有效延缓采后芒果果实的成熟与软化,从而延长果实货架寿命。     

Separation and structure analysis of trisaccharide isomers produced from lactose by Lactobacillus bulgaricus L3 β-galactosidase

Using lactose as the substrate, galacto-oligosaccharides containing β-d-galactose residues were synthesised with β-galactosidase from Lactobacillus bulgaricus L3. The reaction mixture was fermented by yeast cells to consume the monosaccharides and disaccharides, and then it was fully acetylated in the presence of acetic anhydride under I₂ catalysis. Column chromatography of the resulting products, using ethyl acetate: petroleum ether as the eluent, generated two isomers of trisaccharide derivatives (I and II) in gram scale for the first time. Their structure characteristics were investigated by ESI-MS and NMR spectra. They were identified as (2,3,4,6-tetra-O-acetyl-d-galactopyranosyl)-β-(1 → 6)-(2,3,4-tri-O-acetyl-d-galactopyranosyl)-β-(1 → 4)-1,2,3,6-tetra-O-acetyl-α-d-glucopyranose (I) and (2,3,4,6-tetra-O-acetyl-d-galactopyranosyl)-β-(1 → 3)-(2,4,6-tri-O-acetyl-d-galactopyranosyl)-β-(1 → 4)-1,2,3,6-tetra-O-acetyl-α-d-glucopyranose (II), respectively. ESI-MS analysis of both deacetylated products of the two trisaccharide derivatives I and II revealed molecular ion peaks of free trisaccharides, which were structurally identified as Gal-β-(1 → 6)-Gal-β-(1 → 4)-Glc and Gal-β-(1 → 3)-Gal-β-(1 → 4)-Glc, respectively.     

Spirostane,furostane and cholestane saponins from Persian leek with antifungal activity

A phytochemical investigation of the seeds of Persian leek afforded the isolation of two new spirostane glycosides, persicosides A (1) and B (2), four new furostane glycosides, isolated as a couple of inseparable mixture, persicosides C1/C2 (3a/3b) and D1/D2 (4a/4b), one cholestane glycoside, persicoside E (5), together with the furostane glycosides ceposides A1/A2 and C1/C2 (6a/6b and 7a/7b), tropeosides A1/A2 and B1/B2 (8a/8b and 9a/9b), and ascalonicoside A1/A2 (10a/10b), already described in white onion, red Tropea onion, and shallot, respectively. Structure elucidation of the compounds was carried out by comprehensive spectroscopic analyses, including 2D NMR spectroscopy and MS spectrometry, and by chemical evidences. The chemical structure of new compounds were identified as (25S)-spirostan-2α,3β,6β-triol 3-O-[β-d-glucopyranosyl-(1 → 3)] [β-d-xylopyranosyl-(1 → 2)]-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside (1), (25S)-spirostan-2α,3β,6β-triol 3-O-[β-d-xylopyranosyl-(1 → 3)] [α-l-rhamnopyranosyl-(1 → 2)]-β-d-glucopyranosyl-(1 → 4)-O-β-d-galactopyranoside (2), furosta-1β,3β,22ξ,26-tetraol 5-en 1-O-β-d-glucopyranosyl (1 → 3)-β-d-glucopyranosyl (1 → 2)-β-d-galactopyranosyl 26-O-α-l-rhamnopyranosyl (1 → 2)-β-d-galactopyranoside (3a,3b), furosta-2α,3β,22ξ,26-tetraol 3-O-β-d-glucopyranosyl (1 → 3)-β-d-glucopyranosyl (1 → 2)-β-d-galactopyranosyl 26-O-β-d-glucopyranoside (4a,4b), (22S)-cholesta-1β,3β,16β,22β-tetraol 5-en 1-O-α-l-rhamnopyranosyl 16-O-α-l-rhamnopyranosyl (1 → 2)-β-d-galactopyranoside (5).     

Isolation and identification of ent-kaurane-type diterpenoids from Rabdosia serra (MAXIM.) HARA leaf and their inhibitory activities against HepG-2, MCF-7, and HL-60 cell lines

A phytochemical investigation was conducted on Rabdosia serra leaf in this work. A new ent-kaurane-type diterpenoid named 6β,14α-dihydroxy-1α,7β-diacetoxy-7α,20-epoxy-ent-kaur-16-en-15-one, together with 6 known compounds were identified, including parvifolin G, effusanin E, lasiodin, nodosin, β-sitosterol, and stigmasterol. It was the first time that parvifolin G and effusanin E were found in R. serra. The assay of inhibition activity against HepG-2, MCF-7, and HL-60 cell lines indicated that 10 compounds (including rosmarinic acid, methyl rosmarinate and pedalitin which were isolated previously), except parvifolin G and stigmasterol, exhibited cytotoxicity against the tested tumour cells. The tumour inhibitory effects of ent-kaurane-type diterpenoids (except parvifolin G) were more effective than those of sterols and phenolics. Both 6β,14α-dihydroxy-1α,7β-diacetoxy-7α,20-epoxy-ent-kaur-16-en-15-one and lasiodin (IC₅₀ < 5 μM) displayed strong cytotoxicity against the tested tumour cells, indicating the potential for new chemotherapeutic drugs.     

Binding of vitamin A with milk α- and β-caseins

The binding sites of retinol and retinoic acid with milk α- and β-caseins were determined, using constant protein concentration and various retinoid contents. FTIR, UV–visible and fluorescence spectroscopic methods as well as molecular modelling were used to analyse retinol and retinoic acid binding sites, the binding constant and the effect of retinoid complexation on the stability and conformation of caseins. Structural analysis showed that retinoids bind caseins via both hydrophilic and hydrophobic contacts with overall binding constants of K_{retinol-α-caseins} = 1.21 (±0.4) × 10⁵ M⁻¹ and K_{retinol-β-caseins} = 1.11 (±0.5) × 10⁵ M⁻¹ and K_{retinoic acid-α-caseins} = 6.2 (±0.6) × 10⁴ M⁻¹ and K_{retinoic acid-β-caseins} = 6.3 (±0.6) × 10⁴ M⁻¹. The number of bound retinol molecules per protein (n) was 1.5 (±0.1) for α-casein and 1.0 (±0.1) for β-casein, while 1 molecule of retinoic acid was bound in the α- and β-casein complexes. Molecular modelling showed different binding sites for retinol and retinoic acid on α- and β-caseins with more stable complexes formed with α-casein. Retinoid–casein complexation induced minor alterations of protein conformation. Caseins might act as carriers for transportation of retinoids to target molecules.     

Chemical characterisation of anthocyanins in tamarillo (Solanum betaceum Cav.) and Andes berry (Rubus glaucus Benth.) fruits

The anthocyanin composition of tamarillo (Solanum betaceum Cav., red variety) and Andes berry (Rubus glaucus Benth.) was determined by HPLC–PDA and HPLC–ESIMS. From the anthocyanin-rich extracts (AREs), pure compounds (1–7) were obtained by MLCCC (multilayer countercurrent chromatography) and further preparative HPLC, and their unequivocal structures were obtained by 1D and 2D NMR analyses. The new anthocyanin delphinidin 3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside-3′-O-β-d-glucopyranoside, as well as the known cyanidin-3-O-rutinoside, pelargonidin-3-O-rutinoside, and delphinidin-3-O-rutinoside were identified as constituents of tamarillo fruit. Although the anthocyanin composition of Andes berry had been reported before in the literature, the unequivocal structure elucidation of the major compound, cyanidin-3-O-α-l-rhamnopyranosyl-(1 → 6)-O-β-d-xylopyranosyl-(1 → 2)-β-d-glucopyranoside, was achieved for the first time.     

Determination of vitamin C in tropical fruits: A comparative evaluation of methods

Two analytical methods for extracting vitamin C (l-ascorbic and l-dehydroascorbic acids) in tropical fruits [banana, papaya, mango (at three maturity stages) and pineapple] were evaluated. These methods used ion-pair liquid chromatography (LC) for detecting ascorbic acid, but differed in the preparation of the sample (extraction with 3% metaphosphoric acid −8% acetic acid or 0.1% oxalic acid). Results were validated by comparison with ascorbic acid content obtained by the AOAC’s official titrimetric method, by performing a recovery study and by the determination of within-day repeatability and inter-day reproducibility. There were differences in the efficiency of vitamin C extraction related to the fruit matrix and especially to the maturity stage in climacteric fruits. The LC-extraction method using 3% metaphosphoric acid −8% acetic acid shows high mean recoveries (99 ± 6%) for all matrices assayed, while the LC-extraction method with 0.1% oxalic acid proved to be unacceptable in some cases (unripe, half ripe and ripe banana and ripe mango) obtaining mean recoveries of 39.9 ± 9.1% and 72 ± 13% for banana and mango, respectively. The detection limit achieved with the metaphosphoric acid-acetic acid LC-extraction method for ascorbic acid (0.1 mg/l) allowed the determination of this vitamin in fruits analysed with good precision (5.94–12.8%), making its use as a routine analysis method perfectly valid. Recommendations about storage temperature, methods of thawing l-ascorbic acid extracts and the addition of antioxidants to extracts were made.     

Physicochemical changes in fresh-cut wax apple (Syzygium samarangenese [Blume] Merrill & L.M. Perry) during storage

Physicochemical changes, such as peel and flesh colours, total anthocyanin content, browning index, firmness, total soluble solid (TSS), titratable acidity (TA), sugar acid ratio (TSS/TA), antioxidant capacity, total phenolic content and ascorbic acid content, in fresh-cut Taaptimjan wax apple fruit stored at 4 ± 2 °C and 12 ± 2 °C for 7 days were investigated. The skin of fresh-cut fruit stored at 4 ± 2 °C showed higher a^∗ value, chroma and total anthocyanin content and lower hue angle than those stored at 12 ± 2 °C. Lightness (L^∗ value) and whiteness index of the fresh-cut fruit flesh stored at 12 ± 2 °C showed significantly lower than those stored at 4 ± 2 °C which related to an significant increase in browning index. Firmness, total soluble solid, titratable acidity and sugar acid ratio did not significant changes during storage. Antioxidant capacity and total phenolic content increased throughout storage. Ascorbic acid content of the fresh-cut fruit stored at 4 ± 2 °C remained constant throughout storage whilst ascorbic content at 12 ± 2 °C decreased and was lower than that at 4 ± 2 °C. At 4 ± 2 °C antioxidant capacity and ascorbic acid content were higher than that stored at 12 ± 2 °C whilst there was no significant difference in total phenolic content. In conclusion, the reduction of whiteness index and the increase in browning index of fresh-cut wax apple flesh were the key factors affecting its quality and storage at 4 ± 2 °C could reduce the change in the flesh colour and maintained the peel colour and nutritional values of fresh-cut wax apple fruit during storage.     

Treatment of soy milk with Debaryomyces hansenii cells immobilised in alginate

Whole cells of Debaryomyces hansenii UFV-1 were permeabilised with ethanol and immobilised in calcium alginate hydrogel. The optimum pH and temperature for α-galactosidase activities of permeabilised free (PFC) and permeabilised immobilised cells (PIC) were 4.5 and 60 °C; and 4.0 and 70 °C, respectively. PIC α-galactosidase was more stable than that of PFC. The incubation of PIC at 60 and 70 °C promoted an increase in α-galactosidase activity. The α-galactosidase activity was maintained when PIC was used in three repeated batches. The K_m values for PIC and PFC α-galactosidases, with ρNPαGal, were 0.82 mM and 0.30 mM, respectively. Soy milk treatment with PIC for 6 h at 60 °C promoted 100% hydrolysis of raffinose oligosaccharides.     

Purification and characterisation of multiple forms of polygalacturonase from mango (Mangifera indica cv. Dashehari) fruit

Three multiple forms of polygalacturonase (PG) namely PGI, PGII and PGIII were isolated, purified and characterized from ripe mango (Mangifera indica cv. Dashehari) fruit. Native molecular weights of PGI, PGII and PGIII were found to be 120, 105 and 65 kDa, respectively. On SDS–PAGE analysis, PGI was found to be a homodimer of subunit size 60 kDa each while those of PGII and PGIII were found to be heterodimers of 70, 35 and 38, 27 kDa subunit size each, respectively. Three isoforms of PG differed with respect to the effect of pH, metals, reducing agents and their susceptibility towards heat. PG isoforms also differed with respect to the effect of substrate concentration on enzyme activity. PGI and PGIII exhibited inhibition at high substrate concentration while PGII did not. Km for polygalacturonic acid was found to be 0.02% for PGI.