种子脱水耐性与糖的关系

糖类在植物种子中的累积随种子的发育阶段和种子类型不同而不同,并与种子脱水耐性的变化相联系。许多正常性植物种子的发育伴随着某些糖的累积,这些糖的累积已被认为在种子脱水耐性获得中起重要作用。但糖对种子脱水耐性的影响不是单独的,而是与ABA和蛋白质等物质协同作用。种子脱水耐性不仅与糖的种类和含量有关,而且与种子所处的生理状态和发育进程有关。本文综述了种子脱水耐性与糖关系的研究进展。     

兔抗人NESTIN抗血清的制备与鉴定(简报)

巢蛋白(nestin)属Ⅵ类中等纤维蛋白。最初在神经系统发育早期发现有该蛋白的表达。后在正常神经系统中发现仅在未分化的神经前体细胞中有nestin的短暂表达,分化后在神经元和神经胶质细胞中分别被NF(neurofilament,神经丝)和GFAP(Glial fibrillary acidic protein,神经胶质元纤维酸性蛋白)所代替。所以nestin的表达通常被视为神经前体细胞的标志之一。就我们所知,目前商品化nestin的抗体都是鼠抗鼠(克隆Rat401)的抗体,而人鼠之间该蛋白序列的同源性仅为50%左     

Stabilization of Metals in Subsurface by Biopolymers: Laboratory Drainage Flow Studies

Environmental contamination with heavy metals and radionuclides remains a major problem worldwide. The current clean-up methodologies are based on energy-intensive engineering processes, which are disruptive and costly. A new universal technology targeted for the permanent enclosure and fixation of nuclear and other extreme hazardous metallic wastes in subsurface sites is needed. Such technology will be useful in treating contamination at many sites in the U.S., with specific applications to Department of Energy (DOE) sites. Biopolymers are potential tools for such an innovative technology. Biopolymers have repeated sequences, and therefore provide ample opportunity for chemical reactions with metals, soil particles, and other biopolymers. They also have the additional ability of creating cross-linking interpenetrating networks that can encapsulate the contaminants. Based on this concept, in the present work five biopolymers (xanthan, chitosan, polyhydroxy butyrate, guar gum, polyglutamic acid) were investigated for potential use in the stabilization of metals in the subsurface. The effects of these biopolymers (used alone and in combinations) on soil characteristics (permeability, shear strength) and their metal uptake ability have been studied using laboratory drainage flow systems. Biopolymer solutions were run through the experimental sandpack columns, followed by copper solution and leaching agents (distilled water and hydrochloric acid). The permeability and shear strength of sand were evaluated. Copper uptake capacity of each biopolymer and combination of biopolymers was also studied along with subsequent leaching. All biopolymers tested improved sand characteristics (by decreasing permeability and increasing shear strength) and had good metal uptake ability (60–90%) with relatively low leachability (10–22%). While biopolymers used alone were more efficient in metal uptake, the combination of two biopolymers (xanthan and chitosan) had an increasing plugging effect. These results show the potential of using biopolymers in subsurface metal stabilization.     

Synthesis, antifungal activities, and potential detoxification of N-(2,3,4,6-tetra-O-acetyl-β-d-glucopyranosyl)thiocarbamates

A series of N-(2,3,4,6-tetra-O-acetyl-β-d-glucopyranosyl)thiocarbamates were synthesized by the reaction of glucosyl isothiocyanates with monohydric and dihydric alcohols, and acetone oxime, using methods of both normal reaction and microwave-assisted synthesis. Antifungal activities of the title compounds were determined with three kinds of plant pathogenic fungi, Fusarium graminearum, Rhizoctoria cerealis, and Colletotrichum orbiculare. The synthesized glucosyl thiocarbamates easily reacted with HgCl₂ to give novel metal-organic compounds, bis[O-alkyl N-(2,3,4,6-tetra-O-acetyl-β-d-glucopyranosyl)thiocarbamato]mercury, in yields of 80%. This strong affinity of thiocarbamates for mercury showed their potential utility in medical or marine environmental detoxification.     

Is Vitamin E Toxic to Neuron Cells?

Besides acting as potent free radical scavengers, tocopherols and tocotrienols have been known to have non-antioxidant properties such as the involvement of α-tocopherol (αT) in PKC pathway and the anti-cancer properties of γ-tocotrienol (γT3). This study aims to elucidate whether protective effects shown by αT and γT3 in H₂O₂-induced neuron cultures have anti-apoptotic or pro-apoptotic tendency toward the initiation of neuronal apoptosis. H₂O₂ is used to induce apoptosis in primary cerebellar neuron cultures which is attenuated by pretreatment of αT or γT3 at concentrations ≤10 μM. Similar to our previous work, γT3 was found to be neurotoxic at concentrations ≥100 μM, whereas αT showed no neurotoxicity. Cellular uptake of γT3 was higher than that of αT. Treating cells simultaneously with either γT3 or αT and with then H₂O₂ led to higher expression of Bax and Bcl-2 than in neurons exposed to H₂O₂ alone. Analysis of Bcl-2/Bax ratio as ‘survival index’ showed that both pretreatment of γT3 and αT followed by H₂O₂ increase the ‘survival index’ of Bcl-2/Bax ratio compared to H₂O₂-treated cells, while treatment of γT3 alone decrease the ratio compared to unchanged Bcl2/Bax ratio of similar treatment with αT alone. Similar treatment of γT3 decreased p53 expression and activates p38 MAPK phosphorylation, whereas αT did not alter its expression compared to H₂O₂-treated cells. Treating neurons with only γT3 or αT increased the expression of Bax, Bcl-2, p53, and p38 MAPK compared to control with γT3 exerting stronger expression for proteins involved than αT. In conclusion, low doses of γT3 and αT confer neuroprotection to H₂O₂-treated neurons via their antioxidant mechanism but γT3 has stronger pro-apoptosis tendency than αT by activating molecules involved in the neuronal apoptotic pathway in the absence of H₂O₂.     

Proteome alteration of U251 human astrocytoma cell after inhibiting retinoic acid synthesis

Retinoic acid (Ra) is crucial for the patterning and neuronal differentiation in the central nervous system (CNS). Ra deficiency in animals disrupts the motor activities and memory abilities. The molecular mechanisms underlying these behavior abnormalities remain largely unknown. In the current study, we treated the astrocytoma cells with citral, an inhibitor of Ra synthesis. We analyzed the differences in the protein concentrations between the treated and untreated astrocytoma cells by two-dimensional gel electrophoresis (2-DE), Imagemaster software, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In total, 39 of 46 altered protein spots with significant mascot scores were identified representing 36 proteins, that were involved in significantly altered glutamate metabolism, lipid metabolism, mitochrondrial function, and oxidative stress response by Ingenuity Pathway Analysis (IPA). Altered 3-phosphoglycerate dehydrogenase (PHGDH) was also observed in western blot. These data provide some clues for explaining the behavioral changes caused by Ra deficiency, and support the hypothesis that Ra signaling is associated with some symptoms of neurodegenerative disorders and schizophrenia.     

Evidence that TGFA influences risk to cleft lip with/without cleft palate through unconventional genetic mechanisms

This study examined the association between markers in transforming growth factor alpha (TGFA) and isolated, non-syndromic cleft lip with/without palate (CL/P) using a case–parent trio design, considering parent-of-origin effects. We also tested for gene–environmental interaction with common maternal exposures, and for gene–gene interaction using markers in TGFA and another recognized causal gene, IRF6. CL/P case–parent trios from four populations (76 from Maryland, 146 from Taiwan, 35 from Singapore, and 40 from Korea) were genotyped for 17 single nucleotide polymorphisms (SNPs) in TGFA. The transmission disequilibrium test was used to test individual SNPs, and the parent-of-origin likelihood ratio test (PO-LRT) was used to assess parent-of-origin effects. We also screened for possible gene–environment interaction using PBAT, and tested for gene–gene interaction using conditional logistic regression models. When all trios were combined, four SNPs showed significant excess maternal transmission, two of which gave significant PO-LRT values [rs3821261: P = 0.004 and OR(imprinting) = 4.17; and rs3771475: P = 0.027 and OR(imprinting) = 2.44]. Haplotype analysis of these two SNPS also supported excess maternal transmission. We saw intriguing but suggestive evidence of G × E interaction for several SNPs in TGFA when either individual SNPs or haplotypes of adjacent SNPs were considered. Thus, TGFA appears to influence risk of CL/P through unconventional means with an apparent parent-of-origin effect (excess maternal transmission) and possible interaction with maternal exposures.     

Production and characterization of soluble human lysosomal enzyme α-iduronidase with high activity from culture media of transgenic tobacco BY-2 cells

Lysosomal storage diseases (LSDs), that collectively represent over 50 disorders, are amenable to enzyme replacement therapies. However, the current methods used to commercially produce recombinant lysosomal enzymes for this purpose, most commonly Chinese Hamster Ovary cells and human fibroblasts, are prohibitively costly. Plant bioreactors hold great promise for economic production of functional human α-l-iduronidase (hIDUA; glycosaminoglycan α-l-iduronohydrolase; EC 3.2.1.76), the enzyme deficient in the human LSD, Mucopolysaccharidosis I. We have developed and tested an expression system using transgenic tobacco BY-2 cells to produce high amounts of active hIDUA. A plant signal peptide was essential for proper expression and secretion of the 78 kDa glycosylated hIDUA into the cultured media of transgenic BY-2 cells. The yield and activity of the secreted hIDUA from long-term cultures of transgenic BY-2 cell lines were as high as 10 μg/mL media and 53,000 pmol/min/mg proteins, respectively. Thus, this transgenic BY-2 cell line presents an attractive platform for economic production and easy downstream purification of hIDUA for enzyme replacement therapy. Furthermore, this system can be used for the production and purification of other human lysosomal enzymes or pharmaceuticals.     

Plasma proteome analysis of cervical intraepithelial neoplasia and cervical squamous cell carcinoma

Although cervical cancer is preventable with early detection, it remains the second most common malignancy among women. An understanding of how proteins change in their expression during a particular diseased state such as cervical cancer will contribute to an understanding of how the disease develops and progresses. Potentially, it may also lead to the ability to predict the occurrence of the disease. With this in mind, we aimed to identify differentially expressed proteins in the plasma of cervical cancer patients. Plasma from control, cervical intraepithelial neoplasia (CIN) grade 3 and squamous cell carcinoma (SCC) stage IV subjects was resolved by two-dimensional gel electrophoresis and the resulting proteome profiles compared. Differentially expressed protein spots were then identified by mass spectrometry. Eighteen proteins were found to be differentially expressed in the plasma of CIN 3 and SCC stage IV samples when compared with that of controls. Competitive ELISA further validated the expression of cytokeratin 19 and tetranectin. Functional analyses of these differentially expressed proteins will provide further insight into their potential role(s) in cervical cancer-specific monitoring and therapeutics.     

Neurite Outgrowth Effect of 4-O-methylhonokiol by Induction of Neurotrophic Factors Through ERK Activation

Compounds isolated from Magnolia officinalis such as magnolol, honokiol and obovatol exhibit several pharmacological effects on CNS including depressant, anxiolytic and anticonvulsant effects, as well as neuroprotective effects against chemical and heat damages. Recently, honokiol was found to have a neurotrophic effect in fetal rat cortical neurons. In the present study, we show that 4-O-methylhonokiol, a novel compound from Magnolia officinalis, promotes neurite outgrowth in a concentration-dependent manner in rat embryonic neuronal cells. In parallel with the neurite outgrowth activity, the expression of neurite outgrowth marker proteins is also increased by treatment with 4-O-methylhonokiol. We also found that 4-O-methylhonokiol promotes the release of NGF and BDNF into cell culture medium. In addition, lower concentration of 4-O-methylhonokiol (1 and 2 μM) further enhanced neurite outgrowth and expression of neurite outgrowth marker proteins in the presence of NGF (50 ng/ml) or BDNF (10 ng/ml). Subsequently, we found that 4-O-methylhonokiol activates ERK in a concentration-dependent manner. However, the neurite outgrowth activity and the NGF and BDNF release induced by 4-O-methylhonokiol are suppressed by an ERK-specific inhibitor. These results suggest that 4-O-methylhonokiol has the ability to induce neurite outgrowth via the increase of neurotrophic factor levels through ERK activation.