Reversible and irreversible emission of methanethiol and dimethyl disulfide from anaerobically stored broccoli

The reversible and irreversible emission of methanethiol (MT) and dimethyl disulfide (DMDS) from broccoli florets was demonstrated during anaerobic storage at 20 degrees C for up to 24 h. Reversible emission of MT and DMDS was feasible only in broccoli stored for between 0 and 12 h under entirely anaerobic condition. Beyond that, the emission was completely irreversible. This irreversible process was demonstrated through significant reductions in the chlorophyll fluorescence values and rate of carbon dioxide production and significant increase in the membrane permeability of induced broccoli tissues after exposure to air and incubation. Irreversible emission was also demonstrated through significant change in color from the characteristic bright green to olive green as well as the conversion of chlorophyll a to pheophytin a and chlorophyll a' contents of the induced florets after hot-water treatment. These findings suggest that the irreversible emission of MT and DMDS is a function of permanent membrane damage and loss of intracellular compartmentation in the broccoli tissues as a result of the anaerobic induction. The off-odor formation can still be reversed if the affected tissue is only temporarily impaired by anaerobic condition, thereby maintaining the quality of stored broccoli.     

二甲基二硫熏蒸对保护地连作土壤微生物群落的影响

随着保护地高附加值经济作物的连年栽培, 土传病害问题愈发突出, 熏蒸剂也因此得以更广泛的应用。但鉴于熏蒸剂的广谱性, 在杀死有害生物的同时, 不可避免地对非靶标生物产生一定的影响。为明确溴甲烷替代药剂二甲基二硫(dimethyl disulfide, 简称DMDS)熏蒸对土壤微生物群落的影响, 本研究在室内条件下采用BIOLOG 方法, 测定不同浓度DMDS 熏蒸对保护地连作土壤微生物群落的影响。研究结果表明: 不同浓度DMDS(170.00 mg·kg^-1、85.20 mg·kg^-1、42.50 mg·kg^-1、21.30 mg·kg^-1 和10.62 mg·kg^-1)熏蒸处理对镰孢菌属(Fusarium spp.)和疫霉菌属(Phytophthora spp.)的LC₅₀(抑制中浓度)分别为42.08 mg·kg^-1 和115.15 mg·kg^-1。DMDS 熏蒸后恢复培养0 d 取样, 温育120 h 时, 170.00 mg·kg^-1、42.50 mg·kg^-1 和10.62 mg·kg^-1 的DMDS 处理土壤的AWCD 值(平均每孔颜色变化率, average well-color development, AWCD)分别比空白对照升高8.46%、6.02%、19.31%, 表明DMDS 促进了土壤微生物的生长。恢复培养14 d 后, 各处理土壤微生物的AWCD 值恢复至对照水平。多样性指数分析显示, DMDS 熏蒸后恢复培养0 d 时, 土壤微生物群落的Shannon 指数、Simpson指数均高于空白对照, McIntosh 指数与对照无显著性差异; 恢复培养7 d 后, Shannon 指数与Simpson 指数恢复至对照水平。主成分分析结果显示, DMDS 熏蒸后恢复培养0 d 时, 各处理间微生物对碳源的利用方式差异显著, 恢复培养14 d 后, DMDS 对微生物碳源利用方式的影响逐渐减弱, 恢复至对照水平。结果表明, DMDS 熏蒸处理对土壤微生物的生长具有促进作用, 影响了微生物对碳源的利用方式, 但在恢复培养14 d 后, 被干扰的土壤微生物逐渐恢复至对照水平。DMDS 熏蒸处理在有效防控土传病原真菌的同时, 不会对土壤微生物群落产生明显的扰动影响, 对环境较安全。     

Formation of disulfide bonds in acid-induced gels of preheated whey protein isolate

Cold gelation of whey proteins is a two-step process. First, protein aggregates are prepared by a heat treatment of a solution of native proteins in the absence of salt. Second, after cooling of the solution, gelation is induced by lowering the pH at ambient temperature. To demonstrate the additional formation of disulfide bonds during this second step, gelation of whey protein aggregates with and without a thiol-blocking treatment was studied. Modification of reactive thiols on the surface of the aggregates was carried out after the heat-treatment step. To exclude specific effects of the agent itself, different thiol-blocking agents were used. Dynamic light scattering and SDS-agarose gel electrophoresis were used to show that the size of the aggregates was not changed by this modification. The kinetics of gelation as determined by the development of pH and turbidity within the first 8 h of acidification were not affected by blocking thiol groups. During gelation, formation of large, covalently linked, aggregates occurred only in the case of unblocked WPI aggregates, which demonstrates that additional disulfide bonds were formed. Results of permeability and confocal scanning laser microscope measurements did not reveal any differences in the microstructure of networks prepared from treated or untreated whey protein aggregates. However, gel hardness was decreased 10-fold in gels prepared from blocked aggregates. Mixing different amounts of blocked and unblocked aggregates allowed gel hardness to be controlled. It is proposed that the initial microstructure of the gels is primarily determined by the acid-induced noncovalent interactions. The additional covalent disulfide bonds formed during gelation are involved in stabilizing the network and increase gel strength.     

Health-promoting compounds in broccoli as influenced by refrigerated transport and retail sale period

Total aliphatic and indole glucosinolates, phenolic compounds (flavonoids and hydroxycinnamoyl derivatives), and vitamin C contents were evaluated in freshly harvested broccoli (Brassica oleracea L., var. italica, cv. Marathon) inflorescences. These were film-wrapped and stored for 7 days at 1 degrees C to simulate a maximum period of commercial transport and distribution. After cold storage, inflorescences were kept for 3 days at 15 degrees C to simulate a retail sale period. For wrapping, low-density polyethylene (LDPE) of 11 microm thickness was used. Gas composition was about 17% O(2) and 2% CO(2) during cold storage and about 16% O(2) and 3-4% CO(2) during shelf life within packages. The predominant glucosinolates were 4-methylsulfinylbutyl-glucosinolate (glucoraphanin), 3-indolylmethyl-glucosinolate (glucobrassicin), and 1-methoxy-3-indolylmethyl-glucosinolate (neoglucobrassicin). The predominant hydroxycinnamoyl derivatives were identified as 1,2,2'-trisinapoylgentiobiose, 1,2-diferuloylgentiobiose, 1,2'-disinapoyl-2-feruloylgentiobiose, and 3-O-caffeoyl-quinic (neochlorogenic acid). Results showed major losses at the end of both periods, in comparison with broccoli at harvest. Thus, the respective losses, at the end of cold storage and retail periods, were 71-80% of total glucosinolates, 62-59% of total flavonoids, 51-44% of sinapic acid derivatives, and 73-74% caffeoyl-quinic acid derivatives. Slight differences in all compound concentrations between storage and retail sale periods were detected. Distribution and retail periods had minimal effects on vitamin C. Weight loss was monitored at the end of both periods.     

Comparison of isothiocyanate metabolite levels and histone deacetylase activity in human subjects consuming broccoli sprouts or broccoli supplement

Increased consumption of cruciferous vegetables such as broccoli may reduce the risk of various cancers. Myrosinase is required to convert dietary glucosinolates from broccoli into bioactive isothiocyanates. We evaluated isothiocyanate excretion profiles in healthy subjects who consumed broccoli sprouts or broccoli supplement (no myrosinase) with equivalent glucosinolate content. Urinary metabolites of two major isothiocyanates, sulforaphane and erucin, were measured by liquid chromatography coupled with tandem mass spectrometry. Peak excretion of sulforaphane and erucin was higher and occurred sooner in subjects who consumed broccoli sprouts as compared to subjects who consumed the supplement. A subject-dependent shift in the ratio of urinary sulforaphane to erucin metabolites was observed in both groups, indicating conversion of sulforaphane to erucin. Lower histone deacetylase activity was observed in the peripheral blood mononuclear cells only in subjects consuming sprouts. Fresh broccoli sprouts differ from broccoli supplements in regards to excretion of isothiocyanates and bioactivity in human subjects.     

Formation of metal-humic acid complexes by titration and their characterization by differential thermal analysis and infrared spectroscopy

Humic acid (HA) extracted from a Eustis loamy sand (Psammentic Paleudult, Red Yellow Podzolic soil) was flocculated by titration with Al³⁺-, Fe³⁺-, Cu²⁺-, Zn²⁺-, Mn²⁺-, Ba²⁺-, Ca²⁺-, and Mn²⁺-chloride solutions, respectively, to determine possible development of metal-HA complexes, as reported by Flaig et al. (1975), and Tiurin and Kononova (1962). Titration was conducted with HA solutions with an initial pH 11.5 or 7.0. The results indicated that the cations used, except Mg²⁺, yielded insoluble complexes with HA, irrespective of initial pH. After titration, the pH of the metal-HA flocs was 6.0–7.0, which was expected in view of the presence of cation exchange and buffering capacity of HA compounds. More complex formation through electrovalent and covalent bonding by COO^? and phenolic OH groups of the HA molecule was only attained by the use of HA solutions with pH 11.5. On the other hand, less complex formation occurred by the use of HA solutions with an initial pH 7.0, through electrovalent bonding by COO^? groups. Differential thermal analysis (d.t.a.) curves of HA showed shifts in temperatures of the main decomposition peak as a result of flocculation with the different metals. Based on the type of the cations involved, the metal-humic acid flocs could be listed in the following decreasing order of thermal stability: Al³⁺ = Zn²⁺ = Mg²⁺ ≥ HA > Ca²⁺ > Ba²⁺ > Fe³⁺ > Cu²⁺ > Mn²⁺. A systematic relationship could not be found indicating that trivalent ions resulted in the formation of thermally less stable metal-humic acid flocs than divalent ions, as has been reported for HA-metal complexes. Physical mixtures of HA and metal hydroxides exhibited d.t.a. features resembling those of original (nontreated) HA, but not those of the HA-metal flocs.Infrared spectroscopy revealed increased absorption for COO^? vibrations at 1620 and 1400cm^?1 in spectrograms of metal-HA flocs compared to that of original humic acid, a phenomenon explained by many authors to be caused by bonding of the metal ions in hydrated form to the carboxyl or phenolic hydroxyl groups or both of the humic acid molecule. HA-flocs formed from solutions with an initial pH 11.5 had identical i.r. spectra compared with those formed from solutions with an initial pH 7.0.     

褪黑素调控呼吸代谢及抗氧化活性延缓采后青花菜衰老

为探明褪黑素（melatonin,MT）对采后青花菜衰老的调控机理,以鲜切青花菜为试验材料,研究了100 μmol/L MT处理对鲜切青花菜黄化指数、色差、内源褪黑素含量、总叶绿素含量、呼吸途径、能量水平、抗氧化物酶及其同工酶活性的影响。结果表明,贮藏第6 d时CK0、CK1组青花菜的黄化指数分别为1.93、2.08,而MT处理组青花菜在贮藏第7 d的黄化指数仅为1.71,同时MT处理明显抑制了采后青花菜L*、a*、b*值的上升（P<0.05）;随着贮藏时间的延长,青花菜叶绿素含量不断下降,然而至第6 d时,MT组青花菜的叶绿素较CK0、CK1组分别高出18.03%、24.15%。另外,MT处理降低了采后青花菜的总呼吸强度、糖酵解途径、三羧酸循环途径和细胞色素呼吸途径的活性（P<0.05）,提高了其交替途径的活性（P<0.05）,并维持了其较高的能荷水平（P<0.05）;此外,MT处理降低了采后青花菜组织内O2??和H2O2含量的积累（P<0.05）,这可能与其维持组织较高的超氧化物歧化酶、过氧化氢酶、过氧化物酶及其同工酶活性有关（P<0.05）。这些结果表明MT处理可通过调控采后青花菜的呼吸代谢及抗氧化活性而延缓其衰老进程,可为褪黑素应用与青花菜贮藏保鲜提供新的理论支撑。     

Distribution of phenolic acids in different tissues of jujube and their antioxidant activity

Free, esterified, glycosided, and insoluble-bound forms of eight phenolic acids in pulp, seed, and peel of jujube are separated and quantified by high performance liquid chromatography with electrochemical detection (HPLC-ECD). In the whole jujube, p-hydroxybenzoic and cinnamic acids are the most abundant phenolic acids. All quantified phenolic acids are mainly present in jujube peel. Phenolic acids in seed and peel are present in the insoluble-bound form, while, in pulp in the glycosided form, the glycosided and insoluble-bound phenolic acid fractions in jujube pulp represent the highest total phenolic content and the strongest antioxidant activity determined by DPPH and FRAP assays. Our results show that most phenolic compounds with antioxidant activity in different tissues of jujube are present as the glycosided and insoluble-bound forms.     

Formation and inhibition of cholesterol oxidation products during marinating of pig feet

Cholesterol oxidation products (COPs), formed during the heating of cholesterol-rich foods, have been shown to cause cancer and coronary heart disease. The objectives of this study were to develop a GC-MS method for the determination of COPs in pig feet meat, skin, and juice during marinating and to study the formation and inhibition of COPs as affected by the incorporation of soy sauce and sugar. Results showed that an HP-5MS column could provide an adequate separation of cholesterol, 5α-cholestane (internal standard), and seven COPs, including 7α-OH, 7β-OH, 5,6β-OH, 5,6α-OH, triol, 25-OH, and 7-keto, within 15 min with a temperature-programming method. Most COPs in pig feet meat were generated at a larger amount than in pig feet skin and marinating juice over a 24 h heating period at about 100 °C. The Maillard browning index rose with increasing heating time, whereas the pH showed a slight change in marinated juice. Both reducing sugar and free amino acid contributed to the formation of Maillard reaction products. The incorporation of soy sauce and crystal sugar into fresh juice was effective in inhibiting COPs formation in pig feet, skin, and juice over a 30 min preheating period.     

不同穗型小麦品种小花发育成粒对氮肥的响应

为探索不同穗型小麦品种小花发育成粒对氮肥的响应,本试验以大穗型品种‘周麦16’和多穗型品种‘豫麦49’为供试材料,设置不同施氮水平0 kg(N)·hm~(-2)、180 kg(N)·hm~(-2)和360 kg(N)·hm~(-2),观察分析了两个穗型小麦品种小花发育动态模式和结实特性。结果显示,随着生长度日(GDD)的增加,不同氮水平下2品种小花发育动态变化趋势相似,小花分化均符合二次曲线方程模式,退化和败育符合一次线性方程,R2均达显著水平。大穗型品种‘周麦16’小花分化总数显著高于多穗型品种‘豫麦49’,在360 kg(N)·hm~(-2)处理差异极显著;大穗型品种‘周麦16’表现出随施氮量增加,小花分化和退化速率提高,有利于可孕小花的形成、增加结实粒数,最终180 kg(N)·hm~(-2)处理结实粒数显著高于其他处理,较360 kg(N)·hm~(-2)处理平均每穗粒数增加2.04粒;多穗型品种‘豫麦49’尽管分化小花总量不高,但小花退化和可孕小花败育速率较低,在180 kg(N)·hm~(-2)施氮水平表现出高的可孕小花数量和结实粒数,但与高施氮处理相比差异不显著。表明就增加穗粒数而言,两品种均以180 kg(N)·hm~(-2)较为适宜,从最终产量及产量构成分析结果来看,‘豫麦49’表现出与穗粒数相同的结果,而‘周麦16’在高施氮条件下可通过增加成穗数和穗粒数提高产量。     

Kinetics of chlorophyll degradation and color loss in heated broccoli juice

Degradation of chlorophyll in broccoli juice occurred at temperatures exceeding 60 degrees C. Chemical analysis revealed that degradation of chlorophyll a and b to pheophytin a and b, respectively, followed first-order kinetics and that chlorophyll a was more heat sensitive than chlorophyll b. Temperature dependencies of chlorophyll a and b degradation rate constants could be described by Arrhenius equations with activation energies (E(a)) of 71.04 +/- 4.89 and 67.11 +/- 6.82 kJ/mol, respectively. Objective greenness measurements, using the -a value as the physical property, together with a fractional conversion kinetic analysis, indicated that green color degradation followed a two-step process. Kinetic parameters for the first degradation step were in accordance with the kinetic parameters for pheophytinization of the total chlorophyll content, as determined by chemical analysis (E(a) approximately 69 kJ/mol). The second degradation step, that is, the subsequent decomposition of pheophytins, was characterized by an activation energy of 105.49 +/- 4.74 kJ/mol.     

Effect of solvents on in vitro pesticides inhibition of atpase in certain tissues of Labeo rohita

This paper deals with the effect of solvents on the pesticide inhibition of ATPase in the gills and brains of a freshwater teleost Labeo rohita after in vitro treatment with chlordane (an organochlorine pesticide) and Metasystox^R (an organophosphate) pesticide after dissolving in different organic solvents viz. benzene, cyclohexane, cyclopentane, dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMS), and acetone/ethanol (1:1) mixture. The inhibitory effect of chlordane was greatly reduced by cyclohexane, cyclopentane and benzene, to a less extent by DMA, DMF, and DMS and to a lesser extent by acetone/ethanol (1:1) mixture. ATPase inhibition by Metasystox^R showed less effect on the inhibition of ATPase activities. Effect on the inhibition of Na⁺, K⁺-ATPase was observed to be greater than on Mg⁺⁺ ATPase in both tissues, but the activity of both the ATPase in gill homogenate was decreased compared to brain homogenate. Choice of solvents can have profound effects on the response of ATPase to chlorinated hydrocarbon pesticides.     

Formation of natamycin:cyclodextrin inclusion complexes and their characterization

Natamycin is a broad spectrum antimycotic with very low water solubility, which is used to extend the shelf life of shredded cheese products. beta-Cyclodextrin (beta-CD), hydroxypropyl beta-cyclodextrin (HP beta-CD), and gamma-cyclodextrin (gamma-CD) were found to form inclusion complexes with natamycin in aqueous solution. The increase in solubility of natamycin with added beta-CD was observed to be linear (type A(L) phase solubility diagram). The 1:1 stability constant of natamycin:beta-CD complex was estimated from its phase solubility diagram to be 1010 M(-1). The phase solubility diagrams of both gamma-CD and HP beta-CD exhibited negative deviation from linearity (type A(N) diagram) and, therefore, did not allow the estimation of binding constants. The water solubility of natamycin was increased 16-fold, 73-fold, and 152-fold with beta-CD, gamma-CD, and HP beta-CD, respectively. The natamycin:CD inclusion complexes resulted in in vitro antifungal activity nearly equivalent to that of natamycin in its free state.     

Kinetics of the stability of broccoli (Brassica oleracea Cv. Italica) myrosinase and isothiocyanates in broccoli juice during pressure/temperature treatments

The Brassicaceae plant family contains high concentrations of glucosinolates, which can be hydrolyzed by myrosinase yielding products having an anticarcinogenic activity. The pressure and temperature stabilities of endogenous broccoli myrosinase, as well as of the synthetic isothiocyanates sulforaphane and phenylethyl isothiocyanate, were studied in broccoli juice on a kinetic basis. At atmospheric pressure, kinetics of thermal (45-60 degrees C) myrosinase inactivation could be described by a consecutive step model. In contrast, only one phase of myrosinase inactivation was observed at elevated pressure (100-600 MPa) combined with temperatures from 10 up to 60 degrees C, indicating inactivation according to first-order kinetics. An antagonistic effect of pressure (up to 200 MPa) on thermal inactivation (50 degrees C and above) of myrosinase was observed indicating that pressure retarded the thermal inactivation. The kinetic parameters of myrosinase inactivation were described as inactivation rate constants (k values), activation energy (Ea values), and activation volume (Va values). On the basis of the kinetic data, a mathematical model describing the pressure and temperature dependence of myrosinase inactivation rate constants was constructed. The stability of isothiocyanates was studied at atmospheric pressure in the temperature range from 60 to 90 degrees C and at elevated pressures in the combined pressure-temperature range from 600 to 800 MPa and from 30 to 60 degrees C. It was found that isothiocyanates were relatively thermolabile and pressure stable. The kinetics of HP/T isothiocyanate degradation could be adequately described by a first-order kinetic model. The obtained kinetic information can be used for process evaluation and optimization to increase the health effect of Brassicaceae.     

冻融交替对不同施肥水平的黑土中氮磷元素含量的影响

为探究冻融交替对不同施肥水平的黑土中氮磷元素含量变化的影响,对东北地区的黑土进行了模拟冻融处理,并分析其氮磷指标（总氮磷、有效氮磷）的变化。结果表明,冻融交替处理后黑土中全氮的含量会降低,而有效氮和有效磷的含量会提高。单独施加无机肥会加剧冻融交替造成的土壤全氮流失（P<0.05）,5次冻融交替后有效氮含量的增长率降低,有效磷含量的增长率升高。5次冻融交替后,施加有机肥的土壤全氮和全磷含量无显著变化,有效氮和有效磷含量增长率均显著提高（P<0.05）。在寒冷地区冻融条件下的农业生产实践过程中,应适当提高有机肥的施用量,以抑制黑土中氮元素的流失,增强氮磷元素的可用性,保持土壤肥力并降低水体富营养化的风险。