吸附剂及培养基组成对香荚兰细胞悬浮培养产生香兰素的影响

本文报道了培养基的组成及添加吸附剂对香荚兰细胞悬浮培养产生香兰素的影响。结果表明,添加吸附剂活性炭及XAD-2后,香荚兰细胞产生的香兰素含量明显增加,而且活性炭的效果优于XAD-2;香荚兰细胞在全组成的MS培养基中香兰素含量均低于由矿物质盐组成培养基中的含量。可以考虑采用二步培养法来培养香荚兰细胞及产生香兰素。     

Physiology of pepper fruit and the metabolism of antioxidants: chloroplasts,mitochondria and peroxisomes

Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitamins A and C and flavonoids. However, information on the role of these beneficial compounds in the physiology of pepper fruit remains scarce. Recent studies have shown that antioxidants in ripe pepper fruit play a key role in responses to temperature changes, and the redox state at the time of harvest affects the nutritional value for human consumption. In this paper, the role of antioxidant metabolism of pepper fruit during ripening and in the response to low temperature is addressed, paying particular attention to ascorbate, NADPH and the superoxide dismutase enzymatic system. The participation of chloroplasts, mitochondria and peroxisomes in the ripening process is also investigated.Scope and Results Important changes occur at a subcellular level during ripening of pepper fruit. Chloroplasts turn into chromoplasts, with drastic conversion of their metabolism, and the role of the ascorbate–glutathione cycle is essential. In mitochondria from red fruits, higher ascorbate peroxidase (APX) and Mn-SOD activities are involved in avoiding the accumulation of reactive oxygen species in these organelles during ripening. Peroxisomes, whose antioxidant capacity at fruit ripening is substantially affected, display an atypical metabolic pattern during this physiological stage. In spite of these differences observed in the antioxidative metabolism of mitochondria and peroxisomes, proteomic analysis of these organelles, carried out by 2-D electrophoresis and MALDI-TOF/TOF and provided here for the first time, reveals no changes between the antioxidant metabolism from immature (green) and ripe (red) fruits.Conclusions Taken together, the results show that investigation of molecular and enzymatic antioxidants from cell compartments, especially chloroplasts, mitochondria and peroxisomes, is a useful tool to study the physiology of pepper fruit, particularly in the context of expanding their shelf-life after harvest and in maintaining their nutritional value.     

Functional reactivity of WT31 monoclonal antibody with T cell receptor-gamma expressing CD3+4-8- T cells

About 3% of normal peripheral blood T lymphocytes have the phenotype CD3+4-8-. The vast majority of these cells lack the conventional TCR-alpha-, beta complex but express the recently identified TCR-gamma, delta/CD3 receptor complex. These TCR gamma+/CD3+ cells were initially discovered by using as a criterion the lack of reactivity with WT31 mAb. This mAb has been reported to recognize a "framework" epitope on the TCR-alpha, beta/CD3 complex. However, using high concentrations of WT31 mAb, low levels of reactivity with the cell membrane of TCR-gamma+ cells can be observed. This reactivity was significantly increased upon removal of sialic acid residues by neuraminidase. In addition, WT31 mAb is capable to induce lysis by TCR-gamma+ clones. Moreover, immunoprecipitation with WT31 by using cell lysates prepared with the mild detergent digitonin resulted in the isolation of the intact TCR-gamma/CD3 complex. Thus, in contrast to what was previously assumed, WT31 mAb also reacts with a functional epitope present on gamma, delta/CD3 T cells, and therefore lack of reactivity with WT31 mAb is not always a proper hallmark for TCR-gamma-expressing cells.     

Production of Manoalide and Its Analogues by the Sponge <Emphasis Type="Italic">Luffariella variabilis</Emphasis> Is Hardwired

The Great Barrier Reef sponge Luffariella variabilis (Poléjaeff 1884) produces a range of potent anti-inflammatory compounds as its major metabolites. These major metabolites—manoalide monoacetate, manoalide, luffariellin A and seco-manoalide—were monitored temporally and spatially to quantify the potential yield from wild harvest or aquaculture. Production of the major metabolites was hardwired at the population level with little variation in space and time over meters to tens of kilometers in the Palm Islands, Queensland, Australia. Manoalide monoacetate (35 to 70 mg g⁻¹ dry weight of sponge) was consistently the most abundant compound followed by manoalide (15 to 20 mg g⁻¹ dry weight). Luffariellin A and seco-manoalide were 10 to 70 times less abundant and varied between 0 and 3 mg g⁻¹ dry weight. On a larger spatial scale, L. variabilis from Davies Reef and Magnetic Island contained the same rank order and yields of compounds as the Palm Islands, indicating a generality of pattern over at least 100 km. The “hardwiring” of metabolite production at the population level by L. variabilis was also reflected in the lack of any inductive effect on metabolite production. In addition, individually monitored sponges produced fixed ratios of the major metabolites over time (years). However, these ratios varied between individuals, with some individuals consistently producing high levels of manoalide and manoalide monoacetate, providing the potential for selection of high-yielding stocks.     

Co‐benefits,trade‐offs,barriers and policies for greenhouse gas mitigation in the agriculture,forestry and other land use (AFOLU) sector

The agriculture, forestry and other land use (AFOLU) sector is responsible for approximately 25% of anthropogenic GHG emissions mainly from deforestation and agricultural emissions from livestock, soil and nutrient management. Mitigation from the sector is thus extremely important in meeting emission reduction targets. The sector offers a variety of cost‐competitive mitigation options with most analyses indicating a decline in emissions largely due to decreasing deforestation rates. Sustainability criteria are needed to guide development and implementation of AFOLU mitigation measures with particular focus on multifunctional systems that allow the delivery of multiple services from land. It is striking that almost all of the positive and negative impacts, opportunities and barriers are context specific, precluding generic statements about which AFOLU mitigation measures have the greatest promise at a global scale. This finding underlines the importance of considering each mitigation strategy on a case‐by‐case basis, systemic effects when implementing mitigation options on the national scale, and suggests that policies need to be flexible enough to allow such assessments. National and international agricultural and forest (climate) policies have the potential to alter the opportunity costs of specific land uses in ways that increase opportunities or barriers for attaining climate change mitigation goals. Policies governing practices in agriculture and in forest conservation and management need to account for both effective mitigation and adaptation and can help to orient practices in agriculture and in forestry towards global sharing of innovative technologies for the efficient use of land resources. Different policy instruments, especially economic incentives and regulatory approaches, are currently being applied however, for its successful implementation it is critical to understand how land‐use decisions are made and how new social, political and economic forces in the future will influence this process.     

Neuronal and mixed neuronal glial tumors associated to epilepsy. A heterogeneous and related group of tumours

The group of brain tumors with mature components encompasses several pathological entities including: the ganglioneuroma; the gangliocytoma; the ganglioglioma; the desmoplastic ganglioglioma; the neurocitoma and a group of glioneuronal hamartomatous tumorous lesions, such as meningoangiomatosis. The dysembryoplastic neuroepithelial tumor is characterized by the presence of multiple cortical nodules made up of small, oligo-like cells and a myxoid pattern rich in mucopolysaccharides. Mature neuronal cells are frequently detected throughout the tumor. Most of them are associated with microhamartias in the adjacent brain and pharmacoresistant epilepsy. The excellent prognosis of the majority of these tumors and the potential for malignant transformation of the glial component in the ganglioglioma are the two most remarkable findings. Histological signs of anaplasia and greater mitotic and proliferative activities are associated with local recurrences. Atypical neurocytomas occur only exceptionally. Treatment choices are surgical resectioning and, in those cases presenting greater proliferative activity and cytological atypia, postoperative radiotherapy may be recommended. This paper reviews this heterogeneous group of neoplasms and hamartomatous lesions, pointing out presumable transitions among the different types of mixed neuronal and glial brain tumors. A single term of "mixed neuronal-glial tumors" is defended, distinguishing different subgroups of tumors, depending on the predominant cellular component.     

Variation in sclerophylly among Iberian populations of Quercus coccifera L. is associated with genetic differentiation across contrasting environments

Evergreen oaks are an emblematic element of the Mediterranean vegetation and have a leaf phenotype that seems to have remained unchanged since the Miocene. We hypothesise that variation of the sclerophyll phenotype among Iberian populations of Quercus coccifera is partly due to an ulterior process of ecotypic differentiation. We analysed the genetic structure of nine Iberian populations using ISSR fingerprints, and their leaf phenotypes using mean and intracanopy plasticity values of eight morphological (leaf angle, area, spinescence, lobation and specific area) and biochemical traits (VAZ pool, chlorophyll and β-carotene content). Climate and soil were also characterised at the population sites. Significant genetic and phenotypic differences were found among populations and between NE Iberia and the rest of the populations of the peninsula. Mean phenotypes showed a strong and independent correlation with both genetic and geographic distances. Northeastern plants were smaller, less plastic, with smaller, spinier and thicker leaves, a phenotype consistent with the stressful conditions that prevailed in the steppe environments of the refugia within this geographic area during glaciations. These genetic, phenotypic, geographic and environmental patterns are consistent with previously reported palaeoecological and common evidence. Such consistency leads us to conclude that there has been a Quaternary divergence within the sclerophyllous syndrome that was at least partially driven by ecological factors.     

Evaluation of Bottlenecks in the Late Stages of Protein Secretion in Bacillus subtilis

Despite a high capacity for secretion of homologous proteins, the secretion of heterologous proteins by Bacillus subtilis is frequently inefficient. In the present studies, we have investigated and compared bottlenecks in the secretion of four heterologous proteins: Bacillus lichenifomis α-amylase (AmyL), Escherichia coli TEM β-lactamase (Bla), human pancreatic α-amylase (HPA), and a lysozyme-specific single-chain antibody. The same expression and secretion signals were used for all four of these proteins. Notably, all identified bottlenecks relate to late stages in secretion, following translocation of the preproteins across the cytoplasmic membrane. These bottlenecks include processing by signal peptidase, passage through the cell wall, and degradation in the wall and growth medium. Strikingly, all translocated HPA was misfolded, its stability depending on the formation of disulfide bonds. This suggests that the disulfide bond oxidoreductases of B. subtilis cannot form the disulfide bonds in HPA correctly. As the secretion bottlenecks differed for each heterologous protein tested, it is anticipated that the efficient secretion of particular groups of heterologous proteins with the same secretion bottlenecks will require the engineering of specifically optimized host strains.