Steroid control of steroidogenesis in isolated adrenocortical cells: molecular and species specificity

The molecular and species specificity of glucocorticoid suppression of corticosteroidogenesis was investigated in isolated adrenocortical cells. Trypsin-isolated cells from male rat, domestic fowl and bovine adrenal glands were incubated with or without steroidogenic agents and with or without steroids. Glucocorticoids were measured by radioimmunoassay or fluorometric assay after 1-2 h incubation. Glucocorticoids suppressed ACTH-induced steroidogenesis of isolated rat cells with the following relative potencies: corticosterone greater than cortisol = cortisone greater than dexamethasone. The mineralocorticoid, aldosterone did not affect steroidogenesis. Suppression by glucocorticoids was acute (within 1-2 h), and varied directly with the glucocorticoid concentration. Testosterone also suppressed ACTH-induced steroidogenesis. Glucocorticoid-type steroids have equivalent suppressive potencies, thus suggesting that these steroids may induce suppression at least partly by a common mechanism. Although corticosterone caused the greatest suppression, testosterone was more potent. The steroid specificity of suppression of cyclic AMP (cAMP)-induced and ACTH-induced steroidogenesis were similar, suggesting that suppression is not solely the result of interference with ACTH receptor function or the induction of adenylate cyclase activity. Exogenous glucocorticoids also suppressed ACTH-induced steroidogenesis of cells isolated from domestic fowl and beef adrenal glands, thus suggesting that this observed suppression may be a general mechanism of adrenocortical cell autoregulation.     

Shallow lake restoration by nutrient loading reduction—some recent findings and challenges ahead

Shallow lakes respond to nutrient loading reductions. Major findings in a recent multi-lake comparison of data from lakes with long time series revealed: that a new state of equilibrium was typically reached for phosphorus (P) after 10–15 years and for nitrogen (N) after <5–10 years; that the in-lake Total N:Total P and inorganic N:P ratios increased; that the phytoplankton and fish biomass often decreased; that the percentage of piscivores often increased as did the zooplankton:phytoplankton biomass ratio, the contribution of Daphnia to zooplankton biomass, and cladoceran size. This indicates that enhanced resource and predator control often interact during recovery from eutrophication. So far, focus has been directed at reducing external loading of P. However, one experimental study and cross-system analyses of data from many lakes in north temperate lakes indicate that nitrogen may play a more significant role for abundance and species richness of submerged plants than usually anticipated when total phosphorus is moderate high. According to the alternative states hypothesis we should expect ecological resistance to nutrient loading reduction and P hysteresis. We present results suggesting that the two alternative states are less stable than originally anticipated. How global warming affects the water clarity of shallow lakes is debatable. We suggest that water clarity often will decrease due to either enhanced growth of phytoplankton or, if submerged macrophytes are stimulated, by reduced capacity of these plants to maintain clear-water conditions. The latter is supported by a cross-system comparison of lakes in Florida and Denmark. The proportion of small fish might increase and we might see higher aggregation of fish within the vegetation (leading to loss of zooplankton refuges), more annual fish cohorts, more omnivorous feeding by fish and less specialist piscivory. Moreover, lakes may have prolonged growth seasons with a higher risk of long-lasting algal blooms and at places dense floating plant communities. The effects of global warming need to be taken into consideration by lake managers when setting future targets for critical loading, as these may well have to be adjusted in the future. Finally, we highlight some of the future challenges we see in lake restoration research.     

The Functional Roles of the MDM2 Splice Variants P2-MDM2-10 and MDM2-∆5 in Breast Cancer Cells

BACKGROUND: MDM2 is a negative regulator of p53 and is upregulated in numerous human cancers. While different MDM2 splice variants have been observed in both normal tissues and malignant cells, their functions are poorly understood. METHODS: We evaluated the effect of MDM2 splice variants by overexpression in MCF-7 cells and analyses of expression of downstream genes (qPCR and Western blot), subcellular localization (immunofluorescence), cell cycle assays (Nucleocounter3000), apoptosis analysis (Annexin V detection), and induction of senescence (β-galactosidase analysis). RESULTS: In a screen for MDM2 splice variants in MCF-7 breast cancer cells, extended with data from healthy leukocytes, we found P2-MDM2-10 and MDM2-Δ5 to be the splice variants expressed at highest levels. Contrasting MDM2 full-length protein, we found normal tissue expression levels of P2-MDM2-10 and MDM2-Δ5 to be highest in individuals harboring the promoter SNP309TT genotype. While we detected no protein product coded for by MDM2-Δ5, the P2-MDM2-10 variant generated a protein markedly more stable than MDM2-FL. Both splice variants were significantly upregulated in stressed cells (P = 4.3 × 10^?4 and P = 7.1 × 10^?4, respectively). Notably, chemotherapy treatment and overexpression of P2-MDM2-10 or MDM2-Δ5 both lead to increased mRNA levels of the endogenous MDM2-FL (P = .039 and P = .070, respectively) but also the proapoptotic gene PUMA (P = .010 and P = .033, respectively), accompanied by induction of apoptosis and repression of senescence. CONCLUSION: We found P2-MDM2-10 and MDM2-Δ5 to have distinct biological functions in breast cancer cells. GENERAL SIGNIFICANCE: Alternative splicing may influence the oncogenic effects of the MDM2 gene.     

基于近10年红球菌发展的文献计量分析

【背景】近些年,越来越多的研究集中在红球菌上,很少有研究人员对现有文献进行全面回顾。【目的】为了探究国内外红球菌领域的研究热点、前沿和未来发展趋势,以便为后续研究人员提供全面直观的参考。【方法】对近10年发表在Web of Science的红球菌领域论文进行统计分析和文献计量分析。通过VOSviewer文献可视化软件绘制作者标签视图和关键词共现网络图。【结果】全球有关红球菌领域的发文量总体呈逐年上升趋势,发表期刊多为微生物学领域的专科期刊,中国和美国的文章发表数和引用数远超其他国家,红球菌的研究内容也主要集中在生物催化、生物降解和非核糖体肽合成酶等方面。【结论】红球菌在世界范围内越来越受到重视,并逐渐成为研究热点,国家和研究机构应继续加强合作,推动红球菌领域的继续发展。     

2011-2015年度国家自然科学基金微生物学学科项目申请与资助情况分析

本文总结了国家自然科学基金委员会生命科学部微生物学学科2011-2015年度各类项目的申请和资助概况,统计分析了五年来项目依托单位和主要研究方向的变化情况,并从鼓励学科交叉、优化资助格局和加强人才培养的角度对微生物学科资助方向进行展望,以期为微生物学领域的科研工作者提供参考。     

欧文氏菌和棒杆菌的属间融合研究*

研究了用原生质体融合技术获得欧文氏菌和棒杆菌的融合细胞。串联发酵Ｄ－葡萄糖产生２－酮基－Ｌ－古龙酸的第一步发酵菌株欧文氏菌ＳＣＢ２４７经０．８ｍｇ／ｍＬ溶菌酶酶解０．５ｈ后,原生质体的形成率和再生率分别为９９．８％和２７．８％。第二步发酵菌株棒杆菌ＳＣＢ３０５８经预处理后由１．３ｍｇ／ｍＬ溶菌酶酶解２ｈ,原生质体的形成率和再生率分别为９９．５％和５６．３％。用携带氨苄青霉素抗性标记的ＳＣＢ２４７和经热灭活的ＳＣＢ３０５８为亲本,在４０％ＰＥＧ６０００和０．２ｍｏｌ／Ｌ新生磷酸钙等适宜条件下融合,融合频率为３．６×１０~(－６)。在非选择和选择培养基上连续传代十几次后,对融合子的单菌形态、染色结果、菌落形态、色泽、总蛋白量、同工酶、发酵性能等方面与亲本进行了比较。结果表明,融合子确系棒杆菌和欧文氏菌的融合细胞。摇瓶发酵结果显示,所得的３８株稳定的融合子中约４０％能转化葡萄糖为维生素Ｃ前体２－酮基－Ｌ－古龙酸。     

An indicator system to assess ecological integrity of managed forests

Ecological integrity of managed forests includes the ability of an ecosystem to support a community of organisms with a similar species composition and functional organization as found in nearby natural systems. We developed an indicator system for ecological integrity based on simulated natural disturbance and indicator species to test if forest condition and habitat in managed forests are similar to that found or expected in natural systems. We then applied the method in an area of the boreal forest (Ontario, Canada) where the objective of Ontario's strategic forest management planning approach is, in part, to conserve ecological integrity through the emulation of the natural disturbance process. Forest condition controls the supply of habitat to support the diversity of native organisms, and historically in boreal forests the natural disturbance process drove forest condition. We selected indicators of forest condition (landscape pattern and compositional mosaic) and habitat function (occupancy rates for a broad range of forest birds), and applied our assessment system to test whether indicators of forest condition and habitat function reflect outcomes expected if natural disturbance processes were successfully emulated. We collected occupancy data in natural and managed forest disturbance types using autonomous acoustic recorders, applied occupancy/detection modeling to estimate corrected occupancy rates (ψ), and then tested for differences in ψ between disturbance types. Some indicators of forest condition were within the range expected under natural disturbance, but we found relatively less old conifer, more young deciduous and greater edge density in managed forests relative to forests of natural disturbance origin. Most species (11 of 14) occurred with equal ψ in habitat originating from the two disturbance types. Brown creeper (Certhia americana), bay-breasted warbler (Mniotilta varia) and red-eyed vireo (Vireo olivaceus) differed between disturbance types. Brown creeper uses older conifer and occurred at lower rates in managed forest, while red-eyed vireo uses a range of deciduous forest ages, and occurred at higher rates in managed forest. Differences in quantity and/or quality of specific habitat types likely explain the responses. The results suggest what directional changes in the forest pattern and compositional mosaic would improve ecological similarity with natural systems, but also indicate what further research is required. We believe this approach to assessing ecological integrity can be adapted to study the effectiveness of conservation management strategies in other systems, and will contribute to adaptive management approaches and evidence-based policy development.     

Is DNA barcoding child's play? Science education and the utility of DNA barcoding for the discrimination of UK tree species

We present the findings of a DNA barcoding study of the UK tree flora, implemented as part of an innovative, research‐based science education programme called ‘Tree School’. The UK tree flora comprises native and introduced species, and is a taxonomically diverse study group for the exploration of the potential and limitations of DNA barcoding. The children participating in the project collected voucher specimens and generated DNA barcode sequences from trees and shrubs found in the grounds and surrounding woodlands of a residential field centre in Dorset, UK. We assessed the potential of rbcL and matK markers for amplification and DNA sequencing success and for species discrimination among the 67 tree and shrub species included in this study. Although we achieved 100% PCR amplification and sequencing success for rbcL and matK, mononucleotide repeats affected sequence quality in matK for some taxonomic groups (e.g. Rosaceae). Species discrimination success ranged from 65% to 71% using tree‐based methods to 86% using BLASTN. The occurrence of known hybrids (diploid and polyploid) and their progenitors on the study site reduced the overall species discrimination success for both loci. This study demonstrates that, even in a floristic context, rbcL and matK alone are insufficient for the discrimination of UK tree species, especially where taxonomically complex groups are present. From a science education perspective, DNA barcoding represents a compelling and accessible platform for the engagement of non‐experts in ongoing research, providing an opportunity for them to contribute authentic scientific data to an international research campaign.     

Perspectives for ecosystem management based on ecosystem resilience and ecological thresholds against multiple and stochastic disturbances

Ecosystem resilience is the inherent ability to absorb various disturbances and reorganize while undergoing state changes to maintain critical functions. When ecosystem resilience is sufficiently degraded by disturbances, ecosystem is exposed at high risk of shifting from a desirable state to an undesirable state. Ecological thresholds represent the points where even small changes in environmental conditions associated with disturbances lead to switch between ecosystem states. There is a growing body of empirical evidence for such state transitions caused by anthropogenic disturbances in a variety of ecosystems. However, fewer studies addressed the interaction of anthropogenic and natural disturbances that often force an ecosystem to cross a threshold which an anthropogenic disturbance or a natural disturbance alone would not have achieved. This fact highlights how little is known about ecosystem dynamics under uncertainties around multiple and stochastic disturbances. Here, we present two perspectives for providing a predictive scientific basis to the management and conservation of ecosystems against multiple and stochastic disturbances. The first is management of predictable anthropogenic disturbances to maintain a sufficient level of biodiversity for ensuring ecosystem resilience (i.e., resilience-based management). Several biological diversity elements appear to confer ecosystem resilience, such as functional redundancy, response diversity, a dominant species, a foundation species, or a keystone species. The greatest research challenge is to identify key elements of biodiversity conferring ecosystem resilience for each context and to examine how we can manage and conserve them. The second is the identification of ecological thresholds along existing or experimental disturbance gradients. This will facilitate the development of indicators of proximity to thresholds as well as the understanding of threshold mechanisms. The implementation of forewarning indicators will be critical particularly when resilience-based management fails. The ability to detect an ecological threshold along disturbance gradients should therefore be essential to establish a backstop for preventing the threshold from being crossed. These perspectives can take us beyond simply invoking the precautionary principle of conserving biodiversity to a predictive science that informs practical solutions to cope with uncertainties and ecological surprises in a changing world.