Identification of Phenolic Compounds and Determination of Antioxidant Activity in Extracts and Infusions of Salvia Leaves

The influence of harvest period and drying method of Salvia officinalis L. leaves on the content of essential oils, polyphenols and antioxidant properties was investigated. Sage leaves were collected twice during plant blossoming (in June and July) and dried traditionally in natural conditions and at 35 °C. Antioxidant activity was assayed in methanol-acetone extracts and infusions of dried leaves with the use of free radical scavenging activity (DPPH) and ferric reducing antioxidant power (FRAP) technique. Total phenolic content in extracts as well as in infusions was determined by the means of Folin–Ciocalteu method. Based on the LC/MS analysis, the polyphenol compounds present in both extracts and in infusions were identified. The extracts contained more polyphenols and were characterized by higher antioxidant activity than infusions. In the extracts significant amount of ferulic acid was found, which was influenced both by the harvest period and drying method. The amount of ferulic acid found in extracts obtained from the June leaves dried traditionally was three times lower (6.185 μg/g DW) than in extracts from July leaves dried in the same conditions. Harvest period had a significant effect on the essential oils’ content, leaves collected in July contained 15% more oils than those collected in June.     

大鼠颈前皮神经一级传入纤维在脊髓胶状质定位投射的定量分析─FRAP法

目的：研究大鼠颈前皮神经传入纤维在脊髓胶状质（SG）的投射定位。方法：用抗氟化物酸性磷酸酶（FRAP）法和显微测量进行定量研究。结果：颈前皮神经向SG纵向投射主要在C1下三分之一至C4。各节段SG水平向“眉样反应带”所测均值（mm）分别为：0.812、0.816、0.846、0.852，而颈前皮神经向C1～C4SG水平向投射测值（mm）分别在0.402～0.613、0.371～0.662、0.397～0.689、0.525～0.722范围。结论：颈前皮神经投射区从上至下主要居SG中间带外侧和外侧带的内侧。     

P2X3 is expressed by DRG neurons that terminate in inner lamina II

The P2X₃ receptor subunit, a member of the P2X family of ATP-gated ion channels, is almost exclusively localized in sensory neurons. In the present study, we sought to gain insight into the role of P2X₃ and P2X₃-containing neurons in sensory transmission, using immunohistochemical approaches. In rat dorsal root ganglia (DRG), P2X₃-immunoreactivity (-ir) was observed in small- and medium-sized neurons. Approximately 40% of DRG neuronal profiles in normal rats contained P2X₃-ir. In rats that had received neonatal capsaicin treatment, the number of P2X₃-positive neurons was decreased by approximately 70%. Analysis of the colocalization of P2X₃-ir with cytochemical markers of DRG neurons indicated that approximately 94% of the P2X₃-positive neuronal profiles were labelled by isolectin B₄ from Bandeiraea simplicifolia, while only 3% contained substance P-ir, and 7% contained somatostatin-ir. In dorsal horn of rat spinal cord, P2X₃-ir was observed in the inner portion of lamina II and was reduced subsequent to dorsal rhizotomy, as well as subsequent to neonatal capsaicin treatment. Finally, P2X₃-ir accumulated proximal to the site of sciatic nerve ligation, and was seen in nerve fibres in skin and corneal epithelium. In summary, our results suggest that P2X₃ is expressed by a functionally heterogeneous population of BSI-B₄-binding sensory neurons, and is transported into both central and peripheral processes of these neurons.     

大鼠肌皮神经一级传入纤维在脊髓胶状质定位投射的定量分析FRAP法

目的：研究大鼠肌皮神经传入纤维在脊髓胶状质（ＳＧ）的投射定位。方法：用抗氟化物酸性磷酸酶（ＦＲＡＰ）法和显微测量进行定量研究。结果：大鼠肌皮神经在ＳＧ纵向投射主要在Ｃ５～Ｃ６,少数实验动物投向Ｃ７上部。Ｃ５～Ｃ７各节段ＳＧ水平向“眉样反应带”所测均值（ｍｍ）分别为０．８４９、０．９４３、０．９６０,而肌皮神经在Ｃ５～Ｃ７各节段ＳＧ水平向投射所测均值（ｍｍ）分别在０．２５０～０．３８２、０．２８８～０．４２５、０．３０１～０．４０２的范围。结论：肌皮神经在Ｃ５～Ｃ７水平向投射区占据ＳＧ中间带的内侧半。     

翁布中酚性化合物体外抗氧化活性的研究

目的 研究藏药翁布中13种酚性化合物的体外抗氧化活性.方法 采用DPPH自由基清除活性试验、FRAP法、ABTS法和酪氨酸酶活性抑制法评价13种酚性化合物的自由基清除能力及抗氧化活性.结果 没食子酸、3,5-二羟基-4-甲氧基苯甲酸清除DPPH自由基的半数抑制浓度(IC50)低于维生素C;FRAP法测定结果中,化合物阿魏酸、阿魏酸葡糖苷、咖啡酸、对羟基桂皮酸、没食子酸、3,5-二羟基-4-甲氧基苯甲酸的总抗氧化值均高于阳性对照Trolox;ABTS法测定结果中,化合物阿魏酸、松柏醇、阿魏酸葡糖苷、咖啡酸、对羟基桂皮酸、没食子酸、4-羟基-α-甲基苯丙醇、4-羟基-3-甲氧基苯甲酸的总抗氧化值相对于Trolox标准溶液均大于0.01 mmol· L-1;13种酚性化合物对酪氨酸酶均具抑制活性,但IC50均高于阳性对照曲酸.结论 阿魏酸、阿魏酸葡糖苷、咖啡酸、对羟基桂皮酸、没食子酸、3,5-二羟基-4-甲氧基苯甲酸具有较强的抗氧化活性.     

Drosophila under the lens: imaging from chromosomes to whole embryos

Microscopy has been a very powerful tool for Drosophila research since its inception, proving to be essential for the evaluation of mutant phenotypes, the understanding of cellular and tissue physiology, and the illumination of complex biological questions. In this article we review the breadth of this field, making note of some of the seminal papers. We expand on the use of microscopy to study questions related to gene locus and nuclear architecture, presenting new data using fluorescence in-situ hybridization techniques that demonstrate the flexibility of Drosophila chromosomes. Finally, we review the burgeoning use of fluorescence in-vivo imaging methods to yield quantitative information about cellular processes. Electronic Supplementary Material Electronic Supplementary Material is available for this article at and accessible for authorized users.     

Antioxidant capacity,total phenolics and nutritional content in selected ethiopian staple food ingredients

Abstract

The total antioxidant capacity, total phenolics content (TPC) and nutritional content of five types of enset (Enset ventricosum) flour in comparison with four staples (teff [Eragrostis tef], wheat, corn and tapioca) were evaluated. Teff, corn and “amicho” (corm of enset) had the highest ferric reducing antioxidant power (FRAP). The FRAP and TPC of teff (1.8?mmol Trolox equivalence/100?g dry matter (DM) and 123.6?mg gallic acid equivalent/100?g DM, respectively) were over 4-fold larger than the lowest obtained from “bulla” (dehydrated juice of pseudostem of enset). Corn had the lowest IC₅₀ value of 1,1-diphenyl-2-picrylhydrazyl radical scavenging (10.27?mg DM mL^?1). Teff had the highest crude fat content (3.71%) and some mineral profile (P, Mg, Mn and Cu). Enset products had higher fiber, Ca, K, Mg and Mn content as compared to wheat and corn. Ethiopian staple teff has a potential for developing value-added food products with nutritional and health benefits.     

Recent Developments in Effective Antioxidants: The Structure and Antioxidant Properties

Since the last few years, the growing interest in the use of natural and synthetic antioxidants as functional food ingredients and dietary supplements, is observed. The imbalance between the number of antioxidants and free radicals is the cause of oxidative damages of proteins, lipids, and DNA. The aim of the study was the review of recent developments in antioxidants. One of the crucial issues in food technology, medicine, and biotechnology is the excess free radicals reduction to obtain healthy food. The major problem is receiving more effective antioxidants. The study aimed to analyze the properties of efficient antioxidants and a better understanding of the molecular mechanism of antioxidant processes. Our researches and sparing literature data prove that the ligand antioxidant properties complexed by selected metals may significantly affect the free radical neutralization. According to our preliminary observation, this efficiency is improved mainly by the metals of high ion potential, e.g., Fe(III), Cr(III), Ln(III), Y(III). The complexes of delocalized electronic charge are better antioxidants. Experimental literature results of antioxidant assays, such as diphenylpicrylhydrazyl (DPPH) and ferric reducing activity power assay (FRAP), were compared to thermodynamic parameters obtained with computational methods. The mechanisms of free radicals creation were described based on the experimental literature data. Changes in HOMO energy distribution in phenolic acids with an increasing number of hydroxyl groups were observed. The antioxidant properties of flavonoids are strongly dependent on the hydroxyl group position and the catechol moiety. The number of methoxy groups in the phenolic acid molecules influences antioxidant activity. The use of synchrotron techniques in the antioxidants electronic structure analysis was proposed.     

Interferon-gamma inhibits intestinal restitution by preventing gap junction communication between enterocytes

BACKGROUND & AIMS: Necrotizing enterocolitis (NEC) is characterized by interferon-gamma (IFN-gamma) release and inadequate intestinal restitution. Because enterocytes migrate together, mucosal healing may require interenterocyte communication via connexin 43-mediated gap junctions. We hypothesize that enterocyte migration requires interenterocyte communication, that IFN impairs migration by impairing connexin 43, and that impaired healing during NEC is associated with reduced gap junctions. METHODS: NEC was induced in Swiss-Webster or IFN(-/-) mice, and restitution was determined in the presence of the gap junction inhibitor oleamide, or via time-lapse microscopy of IEC-6 cells. Connexin 43 expression, trafficking, and localization were detected in cultured or primary enterocytes or mouse or human intestine by confocal microscopy and (35)S-labeling, and gap junction communication was assessed using live microscopy with oleamide or connexin 43 siRNA. RESULTS: Enterocytes expressed connexin 43 in vitro and in vivo, and exchanged fluorescent dye via gap junctions. Gap junction inhibition significantly reduced enterocyte migration in vitro and in vivo. NEC was associated with IFN release and loss of enterocyte connexin 43 expression. IFN inhibited enterocyte migration by reducing gap junction communication through the dephosphorylation and internalization of connexin 43. Gap junction inhibition significantly increased NEC severity, whereas reversal of the inhibitory effects of IFN on gap junction communication restored enterocyte migration after IFN exposure. Strikingly, IFN(-/-) mice were protected from the development of NEC, and showed restored connexin 43 expression and intestinal restitution. CONCLUSIONS: IFN inhibits enterocyte migration by preventing interenterocyte gap junction communication. Connexin 43 loss may provide insights into the development of NEC, in which restitution is impaired.     

Visualization of the dynamic multimerization of human Cytomegalovirus pp65 in punctuate nuclear foci

The phosphorylated protein pp65 of human Cytomegalovirus (HCMV) is the predominant virion protein and the major tegument constituent. It plays important roles in HCMV infection and virion assembly. Live cell imaging and fluorescence recovery after photobleaching (FRAP) analysis showed that HCMV pp65 accumulated dynamically in punctuate nuclear foci when transiently expressed in mammalian cells. Fluorescence resonance energy transfer (FRET) imaging disclosed that pp65 can self-interact in its localization foci. Yeast two-hybrid assay verified that pp65 is a self-associating protein, and the N-terminal amino acids 14–22 were determined to be essential for pp65 self-association. However, these amino acids were not related to pp65 localization in the specific nuclear foci. The interaction of pp65 and ppUL97 was also studied by FRET microscopy, and the result suggested that there is another signal sequence in pp65, being the ppUL97 phosphorylation site, that is responsible for localization of pp65 in nuclear foci. These results help to understand the function of pp65 in HCMV infection and virion morphogenesis.