利用响应面法优化石榴酒发酵工艺条件

攀枝花的石榴品种繁多，籽粒味甜汁多，为了充分利用这一优势资源，本文进行石榴酒发酵，并对其发酵工艺利用响应面分析法进行优化。以酒精度为指标，考虑发酵温度、二氧化硫添加量、酵母接种量对石榴酒发酵酒精度的影响，然后根据中心组合(Box-Benhnken)原理采用三因素三水平的分析法，依据回归确定各工艺条件的影响因素，以石榴酒发酵酒精度为响应面和等高线，分析各个因素的显著性和交互作用，结果表明优化得到石榴酒发酵的最佳工艺条件为二氧化硫添加量51.3 mg/kg，酵母菌接种量5.33%，发酵温度24.98℃，酒酒精度的理论值为9.58%。通过响应面分析优化发酵工艺，为今后石榴酒发酵的更进一步开发提供一定的理论依据。     

Pomegranate juice moderates anxiety- and depression-like behaviors in AlCl3-treated male mice

PurposeAluminum trichloride (AlCl₃) exposure was proven to encourage some behavioral deficits and eventually induces anxiety and depression in rodents animals. Therefore, this experiment aimed to scout about the effects of pomegranate juice on anxiety- and depression-like behaviors caused by AlCl₃ in male mice.MethodsSix groups of male mice were administrated orally for 35 days by PJ and AlCl3. The control group (G-I) received tap water, while the PJ groups (G-II and G-III) were treated with 20 % and 40 % PJ, respectively. The AlCl₃ group (G-IV) was treated with 400 mg/kg/day of AlCl_3, and the last two groups (G-V and G-VI) were treated with AlCl₃ and 20 % PJ or 40 % PJ, respectively. Then, the open-field (O-F), elevated plus maze (EPM), tail suspension (TS), forced swimming (FS), and light/dark box (L/DB) tests were applied for anxiety- and depression-like behavior studies. In addition, neurotransmitters and oxidative parameters in the brain were evaluated. The plasma cortisol was measured at the end of the experiment.ResultsBehavioral analyses showed that PJ inhibited AlCl₃-induced depressive and anxiogenic effects in the O-F, EPM, TS, FS, L/DB tests. In addition, neurochemical results indicated that PJ at 20 % concentration minimized the AlCl₃ toxicity on dopamine (DOP), serotonin (SER), and acetylcholinesterase (AChE) levels in the for-brain of male mice. Moreover, PJ moderated the AlCl₃ effects by decreasing the level of thiobarbituric acid reactive substances (TBARS), and enhancing catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST) and glutathione (GSH) activities. The plasma cortisol increased in male mice treated with AlCl3 and in a group treated with a high dose of PJ.ConclusionOur results proposed that the anxiety- and depression-like behaviors induced by AlCl₃ exposure in male mice can be ameliorated by PJ treatment, probably through the inhibition of oxidative damage and minimizing the changes in neurotransmitters and hormonal activity.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of the dwarf pomegranate (<Emphasis Type="Italic">Punica granatum</Emphasis> L. var. <Emphasis Type="Italic">nana</Emphasis>)

The dwarf pomegranate (Punica granatum L. var. nana) is a dwarf ornamental plant that has the potential to be the model plant of perennial fruit trees because it bears fruits within 1 year of seedling. We established an Agrobacterium-mediated transformation system for the dwarf pomegranate. Adventitious shoots regenerated from leaf segments were inoculated with A. tumefaciens strain EHA105 harboring the binary vector pBin19-sgfp, which contains neomycin phosphotransferase (npt II) and green fluorescent protein (gfp) gene as a selectable and visual marker, respectively. After co-cultivation, the inoculated adventitious shoots were cut into small pieces to induce regeneration, and then selected on MS medium supplemented with 0.5 μM α-naphthaleneacetic acid (NAA), 5 μM N⁶-benzyladenine (BA), 0.3% gellan gum, 50 mg/l kanamycin, and 10 mg/l meropenem. Putative transformed shoots were regenerated after 6–8 months of selection. PCR and PCR-Southern blot analysis revealed the integration of the transgene into the plant genome. Transformants bloomed and bore fruits within 3 months of being potted, and the inheritance of the transgene was confirmed in T₁ generations. The advantage of the transformation of dwarf pomegranate was shown to be the high transformation rate. The establishment of this transformation system is invaluable for investigating fruit-tree-specific phenomena.     

Survival strategies of pomegranate and almond trees in a fluoride polluted area

In the southern suburb of Sfax (Tunisia), almond and pomegranate trees grow in the vicinity of a phosphate fertilizer-producing factory. In order to reveal strategies adopted by these fruit species to live in restrictive conditions, we investigated the effect of fluoride pollution on the distribution of some elements during the growing season. The results show that calcium and magnesium play an important role in trapping fluoride and delaying the appearance of necroses. Sensitive fruit species, like the almond tree, tend to reduce (F-) toxicity by increasing leaf Mg content, following a decrease in leaf calcium. Since the Mg has multiple roles in the plant metabolism, the MgF affinity seems to be temporary and constitutes, in the long run, a limiting factor for the main physiological processes. Therefore, it appears that resistant species, like pomegranate, have the ability, during fluoride intoxication, not only to assimilate calcium ions but also to maintain Mg concentrations at an adequate level, thus ensuring the cell smooth running. When calcium is no more available, Mg is taken from the chlorophyll molecule as MgF2, and so damage may appear as marginal necroses. By comparing fluoride content of necrotic and healthy leaf slices, it appears that marginal necroses represent typical symptoms of fluoride pollution. By increasing their phosphorus concentration, sensitive fruit species tend to reduce damage intensity, whereas resistant ones tend to avoid F- toxicity.     

石榴皮多酚对妊娠期糖尿病大鼠胰岛素抵抗及氧化应激损伤的影响

摘要 目的：探讨石榴皮多酚（PPPs）对妊娠期糖尿病（GDM）大鼠胰岛素抵抗（IR）及氧化应激损伤的影响。方法：45只孕鼠随机分为对照组、模型组和PPPs组,各15只。模型组和PPPs组孕鼠经腹腔注射45 mg/kg链脲佐菌素制备GDM模型大鼠,对照组经腹腔注射生理盐水。模型制备成功后给予PPPs组大鼠按300 mg/kg PPPs灌胃,对照组和模型组灌胃等体积生理盐水。给药14 d后检测空腹尾静脉血中糖化血红蛋白（HbA1c）、空腹血糖（FBG）、空腹胰岛素（FINS）、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、IL-6、超氧化物歧化酶（SOD）、超敏C反应蛋白（hs-CRP）、丙二醛（MDA）、过氧化氢酶（CAT）、尿酸（UA）、尿素氮（BUN）、血清肌酐（SCr）和尿液β-N-乙酰氨基葡萄糖苷酶（NAG）、肾损伤分子1（KIM-1）水平,计算稳态模型的胰岛素抵抗指数（HOMA-IR）。结果：模型组和PPPs组大鼠血清HbA1c、FBG、FINS、TNF-α、IL-1β、IL-6、hs-CRP、MDA、UA、BUN、SCr和尿液NAG、KIM-1水平以及HOMA-IR显著高于对照组,SOD和CAT活性显著低于对照组（P<0.05）。PPPs组大鼠血清HbA1c、FBG、FINS、TNF-α、IL-1β、IL-6、hs-CRP、MDA、UA、BUN、SCr和尿液NAG、KIM-1水平以及HOMA-IR均显著低于模型组,SOD和CAT活性显著高于对照组（P<0.05）。结论：PPPs能够显著减轻妊娠期糖尿病大鼠胰岛素抵抗、氧化应激水平,改善血糖和肾损伤。     

Proteomic exploration of the impacts of pomegranate fruit juice on the global gene expression of prostate cancer cell

Prostate cancer has been known to be the second highest cause of death in cancer among men. Pomegranate is rich in polyphenols with the potent antioxidant activity and inhibits cell proliferation, invasion, and promotes apoptosis in various cancer cells. This study demonstrated that pomegranate fruit juice could effectively hinder the proliferation of human prostate cancer DU145 cell. The results of apoptotic analyses implicated that fruit juice might trigger the apoptosis in DU145 cells via death receptor signaling and mitochondrial damage pathway. In this study, we exploited 2DE‐based proteomics to compare nine pairs of the proteome maps collected from untreated and treated DU145 cells to identify the differentially expressed proteins. Comparative proteomics indicated that 11 proteins were deregulated in affected DU145 cells with three upregulated and eight downregulated proteins. These dys‐regulated proteins participated in cytoskeletal functions, antiapoptosis, proteasome activity, NF‐κB signaling, cancer cell proliferation, invasion, and angiogenesis. Western immunoblotting were implemented to confirm the deregulated proteins and the downstream signaling proteins. The analytical results of this study help to provide insight into the molecular mechanism of inducing prostate cancer cell apoptosis by pomegranate fruit juice and to develop a novel mechanism‐based chemopreventive strategy for prostate cancer.     

The potential role of pomegranate and its nano-formulations on cerebral neurons in aluminum chloride induced Alzheimer rat model

The oxidative stress leading to degenerative changes in the brain of Alzheimer’s disease (AD) is evident. Our aim was to evaluate the therapeutic and protective effects of pomegranate extract (PE) and pomegranate extract-loaded nanoparticles (PE nano) in an AlCl 3-induced AD rat model. Nanoparticles were synthesized with a PE load of 0.68% w/w, and 70 male Wistar rats were divided into 7 groups: Group I was the control, Group II received PE., Group III received PE nano for 2 weeks, Group IV received AlCl 3 (50 mg/kg) daily orally for 4 weeks, Group V received PE for 2 weeks, Group VI received PE nano for 2 weeks, and Groups V and VI were started after AlCl 3 administration was stopped. Group VII received PE for 2 weeks and was stopped before AlCl 3 was administered. The Results revealed that the discrimination index in the novel object recognition test was the least in AD rat model but increased in cases protected with PE treated with PE nano. Similar results were shown based on calculating the brain weight/body weight percent. The biomarkers of antioxidant activity (catalase, glutathione and total antioxidant activity) in brain homogenate were significantly increased in groups treated with either PE or PE nano. The thiobarbituric acid reactive substance measured to estimate lipid peroxidation was significantly increased in AD rat model and decreased in groups protected with PE or treated with PE nano. Histopathological studies using hematoxylin and eosin, cresyl violet, and silver stains revealed hyaline degeneration, chromatolysis, and hallmarks of AD; neurofibrillary tangles and the senile plaques in brains of AD rat model. Restoration of the histological architecture, Nissl granules, and minimal appearance of hallmarks of AD characterized brains treated with PE or PE nano. In conclusion, PE was more effective as a protectant than a therapeutic measure in alleviating the antioxidant, lipid peroxidative effects and histopathological hallmarks in AD brains. But, the therapeutic PE-loaded nanoparticles increased the efficacy of active components and produced similar results as the protective PE.     

MALDI-TOF mass spectrometry of oligomeric food polyphenols

The structural heterogeneity of polyphenols from cranberries, grape seed extracts, sorghum and pomegranate was characterized by MALDI-TOF MS. Polyphenolics were isolated by liquid chromatography and subjected to MALDI-TOF MS using trans-3-indoleacrylic acid as the matrix. Spectrometric analysis gave information on degree of polymerization, monomeric substitution, and the nature of intermolecular bonds. Cranberry polyflavan-3-ols had variation in interflavan bonds (A- and B-types) and contained polyflavan-3-ols linked to anthocyanins through a CH3-CH bridge. Polygalloyl-polyflavan-3-ols in grape seed extract had large variation in the degree of galloyl substitution. Sorghum polyflavans had structural heterogeneity in glycosylation and hydroxylation. Pomegranate hydrolyzable tannins that correspond to previously described structures were detected, such as punicalagin, but others that correspond to oligomeric ellgitannins in which two to five core glucose units are cross-linked by dehydrodigalloyl and or valoneoyl units were also observed. Results demonstrate that large heterogeneity occurs in degree of polymerization, intermolecular bonds, pattern of hydroxylation, and substitution with monosaccharides and gallic acid.     

A biphasic U-shape effect of cellular oxidative stress on the macrophage anti-oxidant paraoxonase 2 (PON2) enzymatic activity

Expression of macrophage paraoxonase 2 (PON2), a cellular lactonase with anti-oxidant and anti-atherogenic properties, was shown to be upregulated under high oxidative stress. The aim of the present study was to analyze the relationship between the extent of cellular oxidative stress in J774A.1 macrophage and PON2 lactonase activity under various levels of oxidation, obtained by cell incubation with either anti-oxidants or oxidants. PON2 activity exhibited a U-shape response curve. In the oxidative stress range below that of control untreated cells, PON2 activity decreased upon increasing macrophage oxidative state, whereas in the range over that of control untreated cells, PON2 activity increased. The biphasic effect of oxidative stress on macrophage PON2 activity could be related to PON2 inactivation (decreased enzymatic activity) under oxidative stress induction at its low range, whereas at high range of oxidative stress, macrophage anti-oxidant compensatory mechanism up-regulates PON2 (increased protein expression), in order to cope with oxidative burden.