Quantitative analysis of glutamatergic and GABAergic neurons expressing 5-HT(2A) receptors in human and monkey prefrontal cortex

5-hydroxytryptamine (5-HT) or serotonin 2A receptors play an important role in modulation of prefrontal cortex (PFC) activity and have been implicated in the physiopathology of psychiatric disorders. There is no quantitative information on the percentage of glutamatergic and GABAergic cells that express 5-HT(2A) receptors in human and monkey PFC. We have used double in situ hybridization to quantify the mRNA co-localization of 5-HT(2A) receptor with the glutamatergic transporter vesicular glutamate transporter 1, and with the GABAergic marker glutamic acid decarboxylase 65/67 and in parvalbumin and calbindin GABAergic cell populations. Our results show that nearly every glutamatergic cell (86-100%) in layers II-V expressed 5-HT(2A) receptor mRNA in both species. This percentage was lower in layer VI (13-31%). In contrast, not all the GABAergic interneurons (13-46%) expressed 5-HT(2A) receptor mRNA. This receptor was expressed in 45-69% of parvalbumin and in 61-87% of calbindin positive cells. These results indicate that, while the majority of glutamatergic neurons can be sensitive to 5-HT action via 5-HT(2A) receptors, this modulation occurs only in a limited population of GABAergic interneurons and provides new neuroanatomical information about the role played by serotonin through 5-HT(2A) receptors in the PFC and on the sites of action for drugs such as antipsychotics and antidepressants used in treatment of psychiatric disorders.     

夏闲期轮耕对小麦田土壤水分及产量的影响

2007-2010年在宁南旱区研究了夏闲期免耕/深松/免耕(T1)、深松/免耕/深松(T2)、连年翻耕(CT)3种耕作方式对麦田土壤水分及产量的影响.结果表明:经过3年夏闲期T1和T2处理后,农田土壤蓄水效率平均分别较连年翻耕处理提高15.2％和26.5％;T1和T2处理的降水潜在利用率较高,分别达到37.8％和38.5％,降水生产效率平均分别较连年翻耕处理提高9.9％和10.7％.夏闲期轮耕能显著降低休闲期的土壤无效蒸发,有效保蓄小麦生长期的土壤水分.在冬小麦生长前期,T1和T2处理0～200 cm土层土壤水分平均分别较连年翻耕处理增加6.8％和9.4％;在拔节-抽穗-灌浆期,与连年翻耕处理相比,两处理可显著提高0 ～ 200 cm土层土壤蓄水量,对作物产量的贡献率较高.不同轮耕模式在增加作物耗水量的同时也提高了作物产量及水分利用效率,与CT处理相比,3年T1和T2处理作物耗水量平均分别提高5.2％和6.1％,产量分别增加9.9％和10.6％,作物水分生产效率分别提高4.5％和4.3％.相关分析表明,在干旱缺水的宁南地区,冬小麦播种期、拔节-抽穗-灌浆期的土壤蓄水量可显著影响产量,尤其抽穗期的土壤蓄水量对产量的影响更大.     

菜豆属（Phaseolus L.）两种食用作物的核型研究

作者研究了红花菜豆品种大白芸豆和菜豆品种珍珠豆的核型。大白芸豆和珍珠豆的核型公式分别为;２ｎ＝２ｘ＝２２＝１４ｍ（２ＳＡＴ）＋６ｓｍ＋２ｓｔ;２ｎ＝２ｘ＝２２＝６ｍ（２ＳＡＴ）＋１４ｓｍ＋２ｓｔ。这两处品种的核型存在明显差异,但也有相似之处。     

火烧对黔中喀斯特山地马尾松林土壤理化性质的影响

在黔中喀斯特山地马尾松人工次生林内取样分析火烧和对照样地间土壤理化指标的变化,研究了火烧对林地土壤理化性质的影响。结果表明马尾松火烧林地表层土壤毛管孔隙度和总孔隙度升高、最大持水量和最小持水量增加,土壤密度和非毛管孔隙度降低、土壤质量含水量和体积含水量减少;土壤有机质、全N量、全P量、全K量,水解N量、有效P量、速效K量、交换性盐基量和pH值增大,阳离子交换量降低。林火对马尾松林地土壤主要理化指标影响的趋势为或表层土壤影响率大于剖面影响率、或表层土壤影响率小于剖面影响率,不同指标在土壤剖面的变化趋势或增加、或降低,对数或幂函数拟合曲线均达相关显著性水平。火烧和对照样地间的表层土壤理化指标变化主要反映了林火影响,近岩层土壤理化指标变化主要是成土母质在空间上的分异,也受生物的影响。乔木层植株死亡率同表层土壤最大持水量、最小持水量、有机质量和全N量的正相关性显著,同土壤密度的负相关性显著;灌木层植株死亡率同表层土壤密度正相关性显著,同毛管孔隙度、总孔隙度、质量含水量、最大持水量、最小持水量、有机质量、全N量、全P量和速效K量的负相关性达显著或极显著水平;灌木层生物损失量同表层土壤密度和有机质量正相关显著,同速效K量的负相关性显著,枯物层生物损失量同pH值的正相关性显著。火烧马尾松林分平均胸径同表层土壤密度正相关性显著,同毛管孔隙度、总孔隙度、质量含水量、最大持水量、最小持水量和有机质量的负相关性显著。     

Community-wide assessment of protein-interface modeling suggests improvements to design methodology

The CAPRI (Critical Assessment of Predicted Interactions) and CASP (Critical Assessment of protein Structure Prediction) experiments have demonstrated the power of community-wide tests of methodology in assessing the current state of the art and spurring progress in the very challenging areas of protein docking and structure prediction. We sought to bring the power of community-wide experiments to bear on a very challenging protein design problem that provides a complementary but equally fundamental test of current understanding of protein-binding thermodynamics. We have generated a number of designed protein-protein interfaces with very favorable computed binding energies but which do not appear to be formed in experiments, suggesting that there may be important physical chemistry missing in the energy calculations. A total of 28 research groups took up the challenge of determining what is missing: we provided structures of 87 designed complexes and 120 naturally occurring complexes and asked participants to identify energetic contributions and/or structural features that distinguish between the two sets. The community found that electrostatics and solvation terms partially distinguish the designs from the natural complexes, largely due to the nonpolar character of the designed interactions. Beyond this polarity difference, the community found that the designed binding surfaces were, on average, structurally less embedded in the designed monomers, suggesting that backbone conformational rigidity at the designed surface is important for realization of the designed function. These results can be used to improve computational design strategies, but there is still much to be learned; for example, one designed complex, which does form in experiments, was classified by all metrics as a nonbinder.     

覆盖对西北旱地春小麦旗叶光合特性和水分利用的调控

2015—2016年在西北黄土高原半干旱区进行大田定位试验,以‘陇春35号’为试验材料,设全膜覆土穴播(PMS)、全砂覆盖穴播(SM)和露地穴播(CK)3个处理,分析旗叶光合特性、春小麦耗水特性和产量构成因子之间的关系.结果表明: PMS和SM 在0～300 cm土层的土壤贮水量在灌浆前分别较CK提高47.8和31.6 mm,灌浆期均较CK降低15.6 mm.PMS和SM提高春小麦挑旗-抽穗期和扬花-灌浆期的土壤耗水.PMS和SM的叶面积指数分别较CK提高59.0%～73.7%和40.1%～52.7%,叶片SPAD值分别较CK提高3.5%～28.4%和2.9%～23.9%.PMS的光合速率和气孔导度在春小麦挑旗、抽穗、扬花期分别较CK提高23.5%、33.0%、17.7%和32.6%、76.4%、66.9%,灌浆期分别较CK降低26.2%和16.4%;PMS和SM的气孔限制值在抽穗、扬花、灌浆期分别较CK降低14.6%、23.9%、22.3%和25.7%、29.8%、17.4%.叶片瞬时水分利用效率PMS在挑旗期较CK提高57.8%,扬花期降低11.2%.PMS的表观量子效率在抽穗、扬花期分别较SM和CK增加22.6%、18.7%和26.8%、14.3%.PMS和SM春小麦的株高和产量构成因子均显著高于CK,且在干旱年份增幅较大;PMS的产量较CK和SM分别提高36.2%和8.7%,水分利用效率分别提高9.4%和3.4%.因此,PMS和SM提高了小麦灌浆前土壤贮水,加剧了挑旗到抽穗和扬花到灌浆期的耗水,提高了小麦叶片SPAD值和叶面积指数,增强了小麦灌浆前旗叶光合功能,促进 “库”的建成和同化物的转运,实现增产和水分高效利用.PMS在丰水年份的增产潜力和干旱年份的适应能力比SM更强.     

Prognostic molecular markers in pediatric liver disease – Are there any?

Pediatric liver disease (PLD) is a major cause of severe morbidity and prolonged hospitalizations in children. Stratifying patients in terms of prognosis remains challenging. The limited knowledge about molecular mechanisms causing and accompanying PLD remains the main obstacle in a search for reliable prognostic biomarkers. A systematic search of MEDLINE via PubMed and Embase via OVID was conducted on studies published between August 2007 and August 2017. Molecular markers with a prognostic potential in terms of survival, need for liver transplantation or disease progression/regression were selected. In general, identified studies were single center smaller case-control studies or case series with a low level of evidence and a high risk of bias. Only 23 studies comprising 898 patients could be included, mostly focusing on biliary atresia, non-alcoholic fatty liver disease, viral hepatitis, and LT; and markers related to morphogenesis and fibrosis. Furthermore, molecular markers in metabolic pathways and inflammation shown to be relevant, however requiring further validation. Hence, further biological and clinical studies are needed to gain greater molecular insight into PLD.     

A simple system for monitoring biodiversity in protected areas of a developing country

The achievements of initiatives to strengthen biodiversity conservation in developing countries may be difficult to assess, since most countries have no system for monitoring biodiversity. This paper describes a simple and cost-effective, field-based biodiversity monitoring system developed specifically for areas where specialist staff is lacking. We discuss the preliminary lessons learned from protected areas in the Philippines. Whilst the monitoring system aims to identify trends in biodiversity and its uses so as to guide management action, it also promotes the participation of local people in the management, stimulates discussions about conservation amongst stakeholders and builds the capacity of park staff and communities in management skills. In addition, it seeks to provide people with direction regarding the aims of protected areas, and reinforces the consolidation of existing livelihoods through strengthening community-based resource management systems. The field methods are: (1) standardised recording of routine observations, (2) fixed point photographing, (3) line transect survey, and (4) focus group discussion. Both bio-physical and socio-economic data are used and given equal importance. The system can be sustained using locally available resources. The approach is useful in countries embarking on shared management of park resources with local communities, where rural people depend on use of natural ecosystems, and where the economic resources for park management are limited. We hope this paper will encourage other countries to develop their own biodiversity monitoring system, letting its development become a means for capacity building whilst at the same time supporting the creation of ownership.     

High value of short rotation coppice plantations for phytodiversity in rural landscapes

The demand for wood from short rotation coppice (SRC) plantations as a renewable energy source is currently increasing and could affect biodiversity in agricultural areas. The objective was to evaluate the contribution of SRC plantations to phytodiversity in agricultural landscapes assessed as species richness, species–area relationships, Shannon indices, detrended correspondence analysis on species composition, Sørensen similarities, habitat preference proportions, and species proportions found in only one land use. Vegetation surveys were conducted on 12 willow (Salix spp.) and three poplar (Populus spp.) coppice sites as well as on surrounding arable lands, grasslands and forests in central Sweden and northern Germany. SRC plantations were richer in plant species (mean: 30 species per 100 m²) than arable land (10), coniferous forests (13) and mixed forests in Germany (12). Comparing SRC plantations with other land uses, we found lowest similarities in species composition with arable lands, coniferous forests and German mixed forests and highest similarities with marginal grassland strips, grasslands and Swedish mixed forests. Similarity depended on the SRC tree cover: at increased tree cover, SRC plantations became less similar to grasslands but more similar to forests. The SRC plantations were composed of a mixture of grassland (33%), ruderal (24%) and woodland (15%) species. Species abundance in SRC plantations was more heterogeneous than in arable lands. We conclude that SRC plantations form novel habitats leading to different plant species composition compared to conventional land uses. Their landscape‐scale value for phytodiversity changes depending on harvest cycles and over time. As a structural landscape element, SRC plantations contribute positively to phytodiversity in rural areas, especially in land use mosaics where these plantations are admixed to other land uses with dissimilar plant species composition such as arable land, coniferous forest and, at the German sites, also mixed forest.     

Quantifying CD4 receptor protein in two human CD4+ lymphocyte preparations for quantitative flow cytometry

Background

In our previous study that characterized different human CD4+ lymphocyte preparations, it was found that both commercially available cryopreserved peripheral blood mononuclear cells (PBMC) and a commercially available lyophilized PBMC (Cyto-Trol™) preparation fulfilled a set of criteria for serving as biological calibrators for quantitative flow cytometry. However, the biomarker CD4 protein expression level measured for T helper cells from Cyto-Trol was about 16% lower than those for cryopreserved PBMC and fresh whole blood using flow cytometry and mass cytometry. A primary reason was hypothesized to be due to steric interference in anti- CD4 antibody binding to the smaller sized lyophilized control cells.

Method

Targeted multiple reaction monitoring (MRM) mass spectrometry (MS) is used to quantify the copy number of CD4 receptor protein per CD4+ lymphocyte. Scanning electron microscopy (SEM) is utilized to assist searching the underlying reasons for the observed difference in CD4 receptor copy number per cell determined by MRM MS and CD4 expression measured previously by flow cytometry.

Results

The copy number of CD4 receptor proteins on the surface of the CD4+ lymphocyte in cryopreserved PBMCs and in lyophilized control cells is determined to be (1.45 ± 0.09) × 10⁵ and (0.85 ± 0.11) × 10⁵, respectively, averaged over four signature peptides using MRM MS. In comparison with cryopreserved PBMCs, there are more variations in the CD4 copy number in lyophilized control cells determined based on each signature peptide. SEM images of CD4+ lymphocytes from lyophilized control cells are very different when compared to the CD4+ T cells from whole blood and cryopreserved PBMC.

Conclusion

Because of the lyophilization process applied to Cyto-Trol control cells, a lower CD4 density value, defined as the copy number of CD4 receptors per CD4+ lymphocyte, averaged over three different production lots is most likely explained by the loss of the CD4 receptors on damaged and/or broken microvilli where CD4 receptors reside. Steric hindrance of antibody binding and the association of CD4 receptors with other biomolecules likely contribute significantly to the nearly 50% lower CD4 receptor density value for cryopreserved PBMC determined from flow cytometry compared to the value obtained from MRM MS.

Electronic supplementary material

The online version of this article (doi:10.1186/1559-0275-11-43) contains supplementary material, which is available to authorized users.