微波辅助提取荔枝核黄酮类化合物及其抗氧化性研究

研究微波辅助法提取荔枝核黄酮类化合物的工艺,考察了提取溶剂、微波功率、溶剂用量、辐射时间、提取次数等因素对提取的影响。通过正交实验确定最佳的提取参数为:60%乙醇作为提取溶剂,微波功率700 W,料液比1∶25,辐射时间150 s,提取一次。在此优化条件下用微波辅助,黄酮类化合物的得率为6.86%,提取物中黄酮含量达到36.7%。抗氧化性研究表明荔枝核黄酮类化合物有良好的抗氧化活性,能有效清除羟基自由基(OH.)和超氧阴离子自由基(O2-.)。     

荔枝基因组DNA的提取

介绍一种提取荔枝基因组DNA的有效方法,即在传统CTAB法的基础上,于CTAB抽提液中加入1％的PVP。影响荔枝基因组DNA提取质量的因素有3个：第一是取材,第二是磨样与温育时间,第三是防止褐化。     

Evaluation of the in vitro effect of a Lantana camara extract on the labeling of blood constituents of rats with technetium-99m

Blood constituents labeled with technetium-99m (99mTc) have been used in nuclear medicine procedures and drugs are capable to interfere on this labeling. Lantana camara (lantana) has medicinal properties and it has been used in folk medicine. The aim is to verify the effect of a lantana extract on the labeling of blood constituents with 99mTc. Blood of rats was incubated with extract, stannous chloride and 99mTc, as sodium pertechnetate. Plasma (P) and blood cells (BC) were isolated, also precipitated with trichloroacetic acid and soluble (SF) and insoluble fractions (IF) were separated. The % of radioactivity (%ATI) in these samples was calculated. Samples of labeled BC were washed and the %ATI maintained (%ATI-M) in the BC was determined. The results showed that lantana extract decreased significantly (p < 0.05) in the IF-P from 70.24 +/- 2.59 to 11.95 +/- 3.07. This effect was not observed in the BC and IF-BC. The BC-%ATI-M was significantly (p < 0.05) decreased in all concentrations tested when the BC was washed. This fact was not observed in the control. Substances present on the extract should have redoxi action decreasing the concentration of the stannous ion and this condition could justify the effect on the IF-P. The results about the BC-%ATI-M should indicate a possible effect on the transport of ions through the erythrocyte membrane.     

荔枝果皮乙醇提取液抑菌及抗氧化作用的研究(英文)

以乙醇为溶剂的荔枝果皮提取液,用滤纸片法测定其对常见细菌、霉菌的抑菌圈大小,同时研究了不同浓度乙醇的提取液对抑制柑橘青霉的影响,并通过清除DPPH自由基、OH·试验,测定了荔枝果皮的抗氧化效果.结果表明:荔枝果皮的95%乙醇提取液对大肠杆菌、变形杆菌、柑橘青霉均有不同程度的抑制作用,抑菌圈在8.6～18.7 mm之间.其中,柑橘青霉对95%乙醇提取浓度、含量5 mg/mL的荔枝果皮最为敏感,抑菌圈为18.7 mm.不同含量(2.5、5、10、20、40mg/mL)的荔枝果皮提取液DPPH自由基清除率在90%左右,而OH·清除率分别为1.7%、12.9%.78.6%、97.5%和100%.     

测定荔枝核中的游离氨基酸

采用氨基酸自动分析仪测定荔枝核水提取液、醇提取液样品。在醇提取液中游离氨基酸的含量高于水提取液,在两种提取液中共检出了21种游离氨基酸(其中4种为未知氨基酸,6种必需氨基酸);已知游离氨基酸的质量分数为307μg.kg-1(醇提取液)和269μg.kg-1(水提取液),总游离氨基酸的质量分数为500μg.kg-1,必需氨基酸的量占总游离氨基酸量的44%。结论:荔枝核中的游离氨基酸具有利用价值。     

大兴安岭中段森林土壤的黑碳含量及其在不同粒级中的分布

土壤黑碳由于具有生物化学惰性被视为土壤稳定碳库的主要成分.本文量化了大兴安岭中段森林土壤黑碳,分析了黑碳在各粒级内的分布,并探讨了黑碳的稳定机制及其在土壤碳库中的重要性.结果表明: 土壤黑碳表聚现象明显,表层黑碳占总剖面(64 cm)的68.7%,随着土层加深黑碳含量显著降低,但黑碳占有机碳的比例却呈现上升趋势.气候条件影响大兴安岭地区土壤黑碳的分布,相对寒冷和湿润的气候条件提高了土壤固持黑碳的潜力;土壤黑碳在各粒级内所占比例表现为黏粒>粉粒>细砂>粗砂,尽管黏粒中黑碳含量最高,并随土层深度增加而升高,但黏粒中黑碳占有机碳的比例却无明显变化,黑碳/有机碳的升高主要源于细砂与粉粒中黑碳的增加;黑碳的生物化学惰性是表层黑碳的主要稳定机制,而深层的黑碳除了其自身抗性外,黏粒矿物的化学保护发挥着重要作用;黑碳不仅作为稳定碳库的主要成分,在颗粒有机碳组分中仍占相当大的比例,因此黑碳的存在提高了土壤稳定性碳储量与碳汇能力.     

Biocellulose membranes as supports for dermal release of lidocaine

Biocellulose (BC) is a highly pure form of cellulose, produced in the form of a swollen membrane, with several applications in the biomedical area. In this study, the behavior of BC membranes as systems for topical delivery of lidocaine was evaluated. The BC-lidocaine membranes were prepared and characterized in terms of structural and morphological properties. A uniform distribution of the drug inside the BC membranes was observed. In vitro diffusion studies with Franz cells were conducted using human epidermal membranes and showed that the permeation rate of the drug in BC membranes was slightly slower than that obtained with the conventional systems, which was attributed to the establishment of interactions between the lidocaine molecules and the BC membrane, as evidenced by FTIR and NMR analysis. These results indicate that this methodology can be successfully applied for the dermal administration of lidocaine regarding the release profile and ease of application.     

Use of a bacterial cellulose filter for the removal of oil from wastewater

The present study describes the development of a bacterial cellulose (BC) filter for the treatment of oily waters. BC membranes were produced using an alternative medium containing 2.5 % corn steep liquor. Samples of previously purified membranes were characterized and tested as filters for the separation of oil from water (oil concentrations of 10, 150 and 230 ppm). Flow rate, filter diameter and membrane thickness after 6 and 10 days of cultivation were evaluated in a filtration system constructed in polyvinyl chloride. The BC membranes presented satisfactory flexibility, thermal stability and mechanical strength. However, the membrane obtained after 10 days supported 100 % more force than the membrane obtained after 6 days. The experiments revealed 100 % removal of the oil from all emulsions. The filtration flow rate increased proportionally to the filter diameter and decreased from the 6-day membrane to the 10-day membrane. The results of the present study are promising and demonstrate the efficiency, durability and strength of this novel biodegradable, non-toxic material for the treatment of oily waters generated during industrial activities.     

A domain-specific marker for the hepatocyte plasma membrane. III. Isolation of bile canalicular membrane by immunoadsorption

Previous immunolabeling studies (Roman, L.M., and A.L. Hubbard, 1983, J. Cell Biol., 96:1548-1558; Roman, L.M., and A.L. Hubbard, 1984, J. Cell Biol., 98:1488-1496, companion paper) established leucine aminopeptidase (LAP) as a specific marker for the bile canalicular (BC) domain of the rat hepatocyte plasma membrane (PM). In this study, we have isolated membrane from a sonicated PM vesicle fraction using anti-LAP-coated Staphylococcus aureus cells as a solid-phase immunoadsorbent. The extent and specificity of the immunoadsorption were assessed by following the behavior of LAP (the BC marker) and 32P-labeled membrane phospholipids (a uniform membrane marker). The BC fraction obtained was significantly enriched in LAP (yield: greater than 70% of PM-LAP). Alkaline phosphatase, 5'-nucleotidase, and a 110,000-dalton glycoprotein, HA-4, were enriched in the BC fraction to the same extent as LAP (enzyme or antigen/LAP = 1.0). However, alkaline phosphodiesterase I was not enriched to the same degree (enzyme/LAP = 0.5). Contamination of this BC fraction by membrane derived from the sinusoidal domain and endoplasmic reticulum, as determined from the distribution of the asialoglycoprotein receptor and NADH cytochrome c reductase, respectively, was small (less than 13%).     

Molecular mechanism for depolarization-induced modulation of Kv channel closure

Voltage-dependent potassium (Kv) channels provide the repolarizing power that shapes the action potential duration and helps control the firing frequency of neurons. The K⁺ permeation through the channel pore is controlled by an intracellularly located bundle-crossing (BC) gate that communicates with the voltage-sensing domains (VSDs). During prolonged membrane depolarizations, most Kv channels display C-type inactivation that halts K⁺ conduction through constriction of the K⁺ selectivity filter. Besides triggering C-type inactivation, we show that in Shaker and Kv1.2 channels (expressed in Xenopus laevis oocytes), prolonged membrane depolarizations also slow down the kinetics of VSD deactivation and BC gate closure during the subsequent membrane repolarization. Measurements of deactivating gating currents (reporting VSD movement) and ionic currents (BC gate status) showed that the kinetics of both slowed down in two distinct phases with increasing duration of the depolarizing prepulse. The biphasic slowing in VSD deactivation and BC gate closure was strongly correlated in time and magnitude. Simultaneous recordings of ionic currents and fluorescence from a probe tracking VSD movement in Shaker directly demonstrated that both processes were synchronized. Whereas the first slowing originates from a stabilization imposed by BC gate opening, the subsequent slowing reflects the rearrangement of the VSD toward its relaxed state (relaxation). The VSD relaxation was observed in the Ciona intestinalis voltage-sensitive phosphatase and in its isolated VSD. Collectively, our results show that the VSD relaxation is not kinetically related to C-type inactivation and is an intrinsic property of the VSD. We propose VSD relaxation as a general mechanism for depolarization-induced slowing of BC gate closure that may enable Kv1.2 channels to modulate the firing frequency of neurons based on the depolarization history.     

Fatty acids derived from a marine crustacean Diogenes avarus (Heller) and their antiangiogenic activity

An organic extract from a marine crustacean D. avarus was examined for antiangiogenic activity by using the chick embryo chorioallantoic membrane (CAM) assay. The methanol extract (HCM) inhibited angiogenesis in a dose dependent manner. The extract was further fractionated by bioactivity-guided separation to purify the active fractions successively. This resulted in three fractions HCM1, HCM2 and HCM3. The 50% inhibition shown by HCM was 600 ng/disc, HCM1 was 100 ng/disc and of HCM3 was 2.7ng/disc. HCM3 which was separated by column chromatography and showed single spot on TLC was analysed by GLC and showed the presence of saturated and unsaturated fatty acids such as lauric, myristic, palmitic, stearic, oleic, linoleic. The antiangiogenic activity of the fatty acids obtained from a marine crustacean is reported for the first time.     

荔枝胚胎败育与酚类抑制物质的关系

在荔枝 (LitchichinensisSonn .)胚胎败育发生期 ,以系统溶剂法从正常或败育胚珠中初步提取酚类抑制物质 ,通过TLC分离与纯化 ,用GC MS联用仪进一步分离鉴定 ,并以标准品核对。试验首次从荔枝胚珠中分离鉴定出酚类抑制物质对羟基苯甲酸 (p_HBA)。生物活性测定表明 ,p_HBA是一种很强的生长抑制物质。在败育胚珠中其含量及IAA氧化酶活性均显著高于正常胚珠 ,IAA水平则明显低于正常胚珠 (P <0 .0 1)。因此认为 ,p_HBA参与了荔枝胚胎发育的调节 ,高含量的p_HBA是通过促进IAA侧链的氧化并影响促进和抑制生长的物质之间的平衡而导致荔枝胚胎的败育     

Protective effect of bicarbonate against extraction of the extrinsic proteins of the water-oxidizing complex from Photosystem II membrane fragments

A protective effect of bicarbonate (BC) against extraction of the extrinsic proteins, predominantly the Mn-stabilizing protein (PsbO protein), during treatment of Photosystem II (PS II) membrane fragment from pea with 2 M urea, and at low pH (using incubation in 0.2 M glycine-HCl buffer, pH 3.5 or 0.5 M citrate buffer, pH 4.0-4.5) was detected. It was shown that the extraction of the proteins with Mw 24 kDa (PsbP protein) and 18 kDa (PsbQ protein) by the use of highly concentrated solutions of NaCl does not depend on the presence of BC in the medium. An optimal concentration of BC at which it produces the maximum protecting effect was shown to be between 1 mM and 10 mM. The addition of formate did not influence the protein extraction but it reduced the stabilizing effect of BC. Independence of the stabilizing effect on the presence of the functionally active Mn within the water-oxidizing complex indicates that the protecting effect of BC is not related to its interaction with Mn ions. The fact that there is a preferable sensitivity of the PsbO protein to the absence of BC in the medium during all the treatments makes it possible to suggest that either BC interacts directly with the PsbO protein or it binds to some other sites within PS II and this binding facilitates the preservation of the native structure of this protein.     

Effects of preservation media on in vitro maturation of porcine oocytes

The effects of preservation media for ovaries on in vitro maturation of porcine oocytes was studied. The cumulus-oocyte complexes (COCs) obtained from ovaries that had been preserved in three different media at various temperatures for different time intervals were cultured in the M199 maturation medium. The preservation media used were 0.9% saline solution, BCS (Braun-Collins solution) and Dulbecco's phosphate buffered saline solution (PBS). Mature oocytes obtained from the ovaries preserved in three preservation media for 8 h were electrically activated. The activated oocytes were then cultured in the NCSU23 embryo culture medium for 16 h to observe activation, or for 144 h to observe embryo development. It was found that the preservation temperature significantly affected maturation of the porcine oocytes. A preservation temperature of about 25 degrees C showed an optimal maturation rate for a preservation time of 8 h for the three preservation media. Although the preservation temperature was a major factor influencing the maturation rate, different preservation media at 25 degrees C for 8 h also significantly affected the maturation rate, activation rate and embryo development. Among these three preservation media, PBS exhibited the highest cleavage rate indicating that PBS should be a better preservation medium for porcine ovaries at 25 degrees C for 8 h or longer periods.     

Considerations on the flow configuration of membrane chromatography devices for the purification of human basic fibroblast growth factor from crude lysates

Membrane chromatography possesses numerous advantages such as operation at high flow rates, low back pressure, ease of handling and scale up, which make the membrane adsorber process a viable alternative to conventional packed column chromatography. A purification process for the isolation of human recombinant basic fibroblast growth factor (FGF‐2) based on membrane chromatography was investigated using devices with different flow configurations. In the first process, the FGF‐2 capture step was performed with an axial flow device, while the alternative method achieved direct capture of FGF‐2 from unclarified cell lysate with a tangential flow device. In both processes, FGF‐2 purities exceeded 82% and the purified cytokine displayed high biological activity. Binding capacity (BC) from fermentation broth of the axial flow device was 28 mg/mL. This was 50% higher than the BC obtained with the tangential flow device under particle‐free supernatant conditions (18 mg/mL) and 150% higher compared to the BC achieved with unclarified cell lysate (11 mg/mL). While membrane chromatography in tangential flow mode omits clarification and thus reduces the number of stages in the downstream process, it displays lower peak resolution and leads to a lower overall process yield.     

Increased water content in bacterial cellulose synthesized under rotating magnetic fields

The current study describes properties of bacterial cellulose (BC) obtained from Komagataeibacter xylinus cultures exposed to the rotating magnetic field (RMF) of 50 Hz frequency and magnetic induction of 34 mT for controlled time during 6 days of cultivation. The experiments were carried out in the customized RMF exposure system adapted for biological studies. The obtained BC displayed an altered micro-structure, degree of porosity, and water-related parameters in comparison to the non-treated, control BC samples. The observed effects were correlated to the duration and the time of magnetic exposure during K. xylinus cultivation. The most preferred properties in terms of water-related properties were found for BC obtained in the setting, where RMF generator was switched off for the first 72 h of cultivation and switched on for the next 72 h. The described method of BC synthesis may be of special interest for the production of absorbent, antimicrobial-soaked dressings and carrier supports for the immobilization of microorganisms and proteins.     

Direct observation of different surface structures on high-resolution images of native halorhodopsin.

Halorhodopsin (HR) was investigated with atomic force microscopic techniques (AFM) in aqueous solution. Two-dimensional (2D) crystals of HR were obtained by purifying an HR membrane fraction with the same buoyant density as the purple membrane (HR-PM) from the overexpressing strain Halobacterium salinarum D2. The membrane patches of HR were immobilized on mica. Images with a resolution up to 14 A were recorded. Crystals showed an orthogonal structure and the orientation of the molecules showed p42(1)2 symmetry; thus, alternate tetramers are inverted in the membrane. The crystal surface was found to display different structures depending on the imaging force used, indicating that some parts of the HR molecule are more rigid but others more compressible. From samples with single tetramers missing in the crystalline patches dimensions of the unit cell could be determined. Helix-connecting loops in single molecules of halorhodopsin were assigned. The images indicate that the large extracellular BC loop covers the whole molecule and is very flexible.     

Production of bacterial cellulose from Komagataeibacter saccharivorans strain BC1 isolated from rotten green grapes

Abstract

Bacterial cellulose (BC) is one of the prominent biopolymers that has been acquiring attention currently due to its distinctive properties and applications in various fields. The current work presents the isolation of Komagataeibacter saccharivorans strain BC1 isolated from rotten green grapes, followed by biochemical and genotypic characterization, which confirmed that the strain is capable of synthesizing cellulose. Further, production media was designed and certain variables such as carbon, nitrogen sources, pH, and temperature were optimized in order to obtain the maximum concentration of cellulose production. We found mannitol to be the ideal carbon source and yeast extract as the ideal nitrogen source with a highest BC dry yield of 1.81?±?0.25?g/100?mL at pH 5.76 for a week at 30?°C.The charcterization of pellicles by FTIR spectrum depicted similar functional groups present in synthesized BC as that of the commercial cellulose. X-ray diffraction revealed that BC showed 82% crystallinity. Surface morphology of the dried pellicle was studied by SEM image which showed that the BC surface was tightly packed with thin fibers with less porosity. Hence the study demonstrates that the isolates of K.saccharivorans could be used to produce a biopolymer in a short period of time using a modified production medium.     

Preparation of Bacterial Cellulose/Inorganic Gel of Bentonite Composite by In Situ Modification

To evaluate the possibility of Bacterial cellulose/Inorganic Gel of Bentonite (BC/IGB) composite production using in situ method, the BC/IGB composite was successfully produced by in situ modification of BC in both HS medium and corncob hydrolysate. The results showed that the BC/IGB composite obtained in HS medium (one classical medium for BC production) had a higher water holding capacity, but the water retention capacity of the BC/IGB composite obtained in corncob hydrolysate was better. The performance of BC/IGB composite depended on the environment of in situ modification. Using different media showed significant influence on the sugar utilization and BC yield. In addition, BC/IGB composite produced by in situ method was compared with that produced by ex situ method, and the results shows that water holding capacity of BC/IGB composite obtained through in situ method was better. XRD results showed the crystallinity of BC/IGB composite related little to its performance as water absorbent. Overall, in situ modification is appropriate for further production of BC composite and other clay materials.     

Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.