Particulate matter (PM2.5) exposure season-dependently induces neuronal apoptosis and synaptic injuries

Epidemiological studies have shown that particulate matter 2.5 (PM_2.5) not only increases the incidence of cardiopulmonary illnesses but also relates to the development of neurodegenerative diseases. Considering that PM_2.5 is highly heterogeneous with regional disparity and seasonal variation, we investigated whether PM_2.5 exposure induced neuronal apoptosis and synaptic injuries in a season-dependent manner. The results indicated that PM_2.5 altered the expression of apoptosis-related proteins (mainly bax and bcl-2), activated caspase-3 and caused neuronal apoptosis. Additionally, PM_2.5 decreased the levels of synaptic structural protein postsynaptic density (PSD-95) and synaptic functional protein N-methyl-D-aspartate (NMDA) receptor subunit (NR2B) expression. These effects occurred in a season-dependent manner, and PM_2.5 collected from the winter showed the strongest changes. Furthermore, the effect was coupled with the inhibition of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) and phosphorylated cAMP-response element binding protein (p-CREB). Based on the findings, we analyzed the correlations between the chemical composition of PM_2.5 samples and the biological effects, and confirmed that winter PM_2.5 played a major role in causing neuronal apoptosis and synaptic injuries among different season samples.     

Construction of l-Isoleucine Overproducing Strains of Corynebacterium glutamicum

Corynebacterium glutamicum   is used for the industrial production of the amino acids l-glutamate (1×10⁶ tons/year) and l-lysine (300×10³ tons/year). The classical approach to obtain amino acid overproducing strains of C. glutamicum was mutagenesis and then a selection of mutants. In the past 10 years the genetic engineering and amplification of genes have become fascinating methods for studying metabolic pathways in greater detail and for constructing microbial strains with desired genotypes. To obtain l-isoleucine overproducing strains of C. glutamicum we therefore studied the l-isoleucine biosynthesis by overexpression of the various corresponding genes. To enable a flux increase in recombinant strains all genes specific for l-threonine and l-isoleucine biosynthesis were cloned from this bacterium. We demonstratet that amplification of the feedback inhibition insensitive homoserine dehydrogenase and homoserine kinase in a high l-lysine overproducing strain enable the channeling of the carbon flow from the intermediate l-aspartate semialdehyde towards homoserine, resulting in an accumulation of l-threonine. To obtain effective l-isoleucine overproduction a deregulated threonine dehydratase was overexpressed in l-threonine producing strains of C. glutamicum. In this way the l-threonine was converted to l-isoleucine, which was secreted up to 30 g/l into the culture medium. Received: 7 April 1998 / Accepted in revised form: 27 August 1998     

Origin of Metazoa: Sponges as Living Fossils

Geodia cydonium  , which code for proteins. The analyses of their deduced amino acid sequences allowed a molecular biological approach to solve the problem of monophyly of Metazoa. Molecules of the extracellular matrix/basal lamina, with the integrin receptor, fibronectin, and galectin as prominent examples, cell-surface receptors (tyrosine kinase receptor), elements of sensory systems (crystallin, metabotropic glutamate receptor), and homologs/modules of an immune system (immunoglobulin like molecules, scavenger receptor cysteine-rich, and short consensus repeats, rhesus system) classify the Porifera as true Metazoa. As living fossils, provided with simple, primordial molecules allowing cell-cell and cell-matrix adhesion as well as processes of signal transduction as known in a more complex manner from higher Metazoa, they also show peculiarities not known in other metazoan phyla. Tissues of sponges are rich in telomerase activity, suggesting a high plasticity in the determination of cell lineages. It is concluded that molecular biological studies with sponges as model will not only help to understand the evolution of Protoctista to Metazoa but also the complex, hierarchial regulatory network of cells in higher Metazoa.     

Cardiac Contractility: How Calcium Activates the Myofilaments

2+   concentrations tropomyosin is located at the edge of the thin filament, thereby interfering with the formation of strong actin-myosin linkages (blocked state). An increase in Ca²⁺ activity causes an azimuthal shift of tropomyosin around the filament (by about 30°), thereby increasing the probability of low-force crossbridge interaction, a process which by cooperative effects induces further tropomyosin movement (by an additional 10°) which results in the open state of the filament characterized by forceful crossbridge interaction. (This mechanism may be analogous to that in ligand-gated ion channels, where ligand binding increases the open probability of the pore.) The extent of activation then depends on the free Ca²⁺ concentration and on the calcium sensitivity of the thin filament that may be affected by protein phosphorylation, crossbridge attachment, the troponin isoform composition of the filament, and novel calcium-sensitizing drugs that act on the contractile or regulatory proteins and thus increase the force of the heart.     

Pasture management under goat grazing on Canary Islands

Tenerife pastures in the Teno plateau are under a goat grazing management system, and the number of goats has increased by 70% in the past 10 years. This plant community was sampled during 4 years (1992, 1993, 1994 and 1999) over a 10-year period, using ten 0.25 m² plots to reveal changes in species composition, soil nutrient content, and biomass nutrient content over time and relate these variables to changes in management and understand aspects of the dynamics of these pastures.The results revealed no changes in biomass nutrient composition over the sampling period; however, biomass protein content was related to changes in species composition. Soil phosphorus content increased in the last sampling year and was related to an average species richness decrease in the plots. Results suggest that the increase in soil phosphorus and the decrease in species richness are related to the increase in goat grazing intensity. At the end of the studied period, species with a higher protein content increased in importance in the species composition. The restoration of pastures degraded by agriculture or overgrazing is required to provide a larger area for grazing and to ensure sustainable management of these protected areas.     

Calcium

Calcium plays a pivotal role in cellular signal transduction. Precise control and regulation of cytoplasmic free ionic calcium concentration is a prerequisite for intracellular calcium-mediated physiology. Calmodulin is the ubiquitous calcium receptor protein which conveys the information to the calcium-dependent enzymes. Protein kinase C and receptor-stimulated phosphatidylinositol metabolisms are components of another regulatory mechanism where calcium plays an important regulatory role, too. Oncogene products have been found to possess enzymatic activities involved in phosphatidylinositol metabolism.     

环境内分泌干扰物促进乳腺细胞增殖的分子机制研究进展

乳腺癌是女性中常见的癌症,EDCs（environmental endocrine disrupting chemicals,环境内分泌干扰物）具有雌激素活性,能促进乳腺细胞的增殖,明确EDCs在乳腺癌细胞增殖中的作用和机制,有助于乳腺癌的预防.在归纳整理国内外相关研究基础上,总结了EDCs通过激活雌激素受体信号通路、抑制细胞凋亡和改变细胞周期等方面促进乳腺细胞的增殖,并对其分子机制进行了全面综述.EDCs促进乳腺细胞增殖的分子机制有:①EDCs通过与雌激素核受体（nERs）结合形成二聚体,改变受体构象,并结合到靶基因雌激素响应元件（ERE）上,进而调控增殖相关靶基因转录和蛋白表达;②EDCs也可通过与雌激素膜受体（mERs）结合,激活快速非基因组信号通路,改变靶蛋白活性或靶基因表达;③EDCs也可通过上调凋亡抑制蛋白Bcl-2和Bcl-xL的表达,抑制细胞凋亡;④EDCs还可通过上调细胞周期蛋白（cyclin）表达、提高周期依赖性蛋白激酶（CDKs）活性,加快细胞周期;⑤EDCs通过基因组、非基因组信号途径的交互,上调cyclin D和c-Myc表达,再通过CDK磷酸化细胞周期调节因子Rb蛋白,释放转录因子E2Fs,加速细胞从G₁期向S期的转变,促进细胞增殖.建议今后从与内源激素交互、影响雌激素E₂合成以及低剂量联合作用3个方面来探讨EDCs对乳腺细胞的增殖机制,以期为乳腺致癌物的筛查、乳腺癌预防和治疗以及乳腺细胞增殖机制的深入研究提供参考.      

Low Pathogenic Avian Influenza Viruses (H3N8, H5N6): In Vitro Influence of <Emphasis Type="SmallCaps">d</Emphasis>,<Emphasis Type="SmallCaps">l</Emphasis>-Lactic Acid and Sodium Chloride on Infectivity and Virus Persistence in Short Fermented Raw Poultry Sausage

Since highly pathogenic avian influenza virus H5N1 emerged in 1997, avian influenza is considered one of the most important infectious diseases globally. In respect of virus transmission to humans, the consumption of raw poultry products remains of serious concern. In this study, data about survival time and inactivation kinetics of two low pathogenic avian influenza virus (AIV) strains (H3N8, H5N6) in short fermented raw sausage were obtained. In addition, the impact of the preserving factors d,l-lactic acid and sodium chloride on virus infectivity was evaluated through in vitro studies. Virus infectivity was confirmed in embryonated chicken eggs. Inactivation of H3N8 was seen in d,l-lactic acid solutions (0.15 and 0.20%, pH 4.40–4.70 and pH 3.80–3.91) at both temperatures (20 vs. 4°C) during 3 days of exposure. However, infectious virus particles could still be detected after exposure to 0.1% d,l-lactic acid (pH 5.80–5.99). In all NaCl solutions (2, 6 and 12% w/v), infectivity of the H3N8 strain decreased steadily but reduction of the virus titre increased significantly with higher temperature. In raw sausages, decline in virus titre was observed for both strains during ripening and storage. Thereby, decline of virus infectivity was dependent on time and temperature with a more marked effect at higher temperatures (22 vs. 7°C). At refrigeration (7°C), both viruses maintained infectivity over 14 days. Results indicate that appropriate processing of short fermented raw poultry sausage is likely to reduce risk of virus exposure due to adequate inactivation of AIV during ripening and storage.     

A putative octopamine/tyramine receptor mediating appetite in a hungry fly

In the blowfly Phormia regina, experience of simultaneous feeding with d-limonene exposure inhibits proboscis extension reflex (PER) due to decreased tyramine (TA) titer in the brain. To elucidate the molecular mechanism of TA signaling pathway related to the associated feeding behavior, we cloned cDNA encoding the octopamine/TA receptor (PregOAR/TAR). The deduced protein is composed of 607 amino acid residues and has 7 predicted transmembrane domains. Based on homology and phylogenetic analyses, this protein belongs to the OAR/TAR family. The PregOAR/TAR was mainly expressed in head, with low levels of expression in other tissues at adult stages. Gene expression profile is in agreement with a plethora of functions ascribed to TA in various insect tissues. The immunolabeled cell bodies and processes were localized in the medial protocerebrum, outer layer of lobula, antennal lobe, and subesophageal ganglion. These results suggest that decrease of TA level in the brain likely affects neurons expressing PregOAR/TAR, causing mediation of the sensitivity in the sensillum and/or output of motor neurons for PER.     

Photoperiod affects daily torpor and tissue fatty acid composition in deer mice

Photoperiod and dietary lipids both influence thermal physiology and the pattern of torpor of heterothermic mammals. The aim of the present study was to test the hypothesis that photoperiod-induced physiological changes are linked to differences in tissue fatty acid composition of deer mice, Peromyscus maniculatus (∼18-g body mass). Deer mice were acclimated for >8 weeks to one of three photoperiods (LD, light/dark): LD 8:16 (short photoperiod), LD 12:12 (equinox photoperiod), and LD 16:8 (long photoperiod). Deer mice under short and equinox photoperiods showed a greater occurrence of torpor than those under long photoperiods (71, 70, and 14%, respectively). The duration of torpor bouts was longest in deer mice under short photoperiod (9.3 ± 2.6 h), intermediate under equinox photoperiod (5.1 ± 0.3 h), and shortest under long photoperiod (3.7 ± 0.6 h). Physiological differences in torpor use were associated with significant alterations of fatty acid composition in ∼50% of the major fatty acids from leg muscle total lipids, whereas white adipose tissue fatty acid composition showed fewer changes. Our results provide the first evidence that physiological changes due to photoperiod exposure do result in changes in lipid composition in the muscle tissue of deer mice and suggest that these may play a role in survival of low body temperature and metabolic rate during torpor, thus, enhancing favourable energy balance over the course of the winter.     

Small-scale response of plant species to land-use intensification

Plant communities are affected by land-use and landscape heterogeneity and can be used as indicators of environmental change. At small-scale, species composition and species richness of plant communities are influenced by local environment and by diaspores from the surroundings. Thus, they reflect the influence of both land-use type and land-use diversity. Plant community composition was studied along a gradient of agricultural disturbance in the Morvan Regional Natural Park (Burgundy, France). Six landscape units of 1 km² were selected along a range of increasing land-use intensity. Sixteen 0.2 m² sampling plots per unit were selected according to a grid-based design to estimate the percent cover of all plant species. Pattern analysis showed that local species richness increased from woodland to crop to grassland, and also increased with land-use diversity. Local plant biodiversity was maximized under intermediate disturbance intensity and minimized at low (woodland) and high (crop) disturbance levels.     

典型酚类污染物内分泌干扰效应研究——对斑马鱼发育及核受体介导基因调控的分子影响

以斑马鱼为研究对象,观察2,4,6-三氯酚（2,4,6-trichlorophenol）、4-氯-3-甲酚（4-chloro-3-methylphenol）和4-硝基苯酚（4-nitrophenol）3种酚类污染物对斑马鱼胚胎发育早期阶段形态学指标的影响,借助q-RT-PCR方法对斑马鱼胚胎分别暴露于0.1,0.5和2.5mg/L至120hpf幼鱼8个重要受体（雄激素受体（AR）、甲状腺激素受体（TR）、芳香烃受体（AhR）、雌激素受体（ER）、糖皮质激素受体（GR）、孕烷X受体（PXR）、盐皮质激素受体（MR）、过氧化物酶体增殖物激活受体（PPAR））相关基因的转录水平进行了研究;进一步采用分子对接和分子动力学模拟的方法研究污染物与筛选出的影响受体（ER、AhR、PXR、GR以及MR）之间的相互作用过程.结果表明,在5和10mg/L的浓度下3种酚类污染物都显著影响斑马鱼的发育,造成脊柱弯曲、心包水肿等不良影响;基因结果显示2,4,6-三氯酚暴露导致ER的基因下调,雌激素受体α（er1）基因在2.5mg/L浓度下下调2.1倍.4-氯-3-甲酚显著影响ER、AhR以及PXR的相关基因,表现为在0.5和2.5mg/L暴露er1基因显著下调1.8和2.2倍,ahr2和pxr基因在0.1和2.5mg/L的浓度下分别下调1.7和1.8倍和上调3.5和3.2倍.4-硝基苯酚暴露使GR、MR核心受体基因发生调控变化,糖皮质激素受体基因（gr）在低中浓度下分别显著上调2.0和2.1倍,最高浓度下下调1.9倍.盐皮质激素受体基因（mr）在中高浓度下分别下调1.9和2.1倍,在最低浓度下显著上调1.7倍;分子计算结果显示它们与相关受体通过疏水和氢键等相互作用而稳定结合,且均方根偏差（RMSD）在5ns后稳定.这3个典型酚类污染物主要通过斑马鱼ER、AhR、PXR、GR、MR受体介导产生内分泌干扰效应并影响其早期生长发育.     

Effect of dung deposition on small-scale patch structure and seasonal vegetation dynamics in mountain pastures

Cattle activity greatly influences plant species composition and biomass production of grassland ecosystems. Dung deposition by cattle together with grazing and trampling can be considered as one of the important factors driving vegetation dynamics in pastures. The objective of this study was to investigate at 10-cm and 1-month resolution the plant community dynamics induced by dung deposition in two plant communities (a mesotrophic and an oligotrophic grassland) in a pasture of the Swiss Jura Mountains. Vegetation was sampled four or three times during the vegetation period in contiguous 10 cm × 10 cm quadrats from the centre of the dung pat to a distance of 60 cm. A lower grazing intensity near the dung pat was recorded for all observation periods. In the mesotrophic grassland the canopy was higher near the dung pat already one week after dung deposition. Vegetation around dung pats was submitted to two opposite fertilizing and grazing gradients, which induced changes in vegetation texture and structure at fine scale and short term. We observed a positive rank correlation between species turnover and distance to the dung for both communities, suggesting a seasonal stabilizing effect of dung on the plant composition of their direct surroundings (0–10 cm) likely due to cattle avoidance. Since dung pats are dropped every year in different locations, they create in the pasture a shifting mosaic of nutrient availability and grazing intensity inducing at seasonal scale micro-successions in plant communities.     

Ancient paddy soils from the Neolithic age in China’s Yangtze River Delta

Identifying prehistoric irrigated rice fields and characterizing the beginning of paddy soil development are important for a better understanding of human development and agricultural history. In 2003, paddy soils and irrigated rice fields buried at a depth of 100–130 cm were excavated at Chuo-dun-shan in the Yangtze River Delta, close to Suzhou, China. The fields of sizes between 1.4 and 16 m² were surrounded with ridges that were connected to ditches/ponds via outlets to control the water level within the fields. Many carbonized and partly carbonized rice grains with an age of 3,903 B.C.(measured ¹⁴C age 5,129±45 a BP) were recovered. The surface layers of these buried paddy fields showed a high content of soil organic matter and a considerable high density of rice opals. The latter were identified to derive from Oryza spp. Solid-state ¹³C nuclear magnetic resonance spectroscopy revealed aromatic carbon (C) as the predominant organic C form in the fossil surface layer. This is expected, if the major source represents burnt rice and straw. In summary, our data are in agreement with new evidences indicating that in China, paddy soils and irrigated rice cultivation were initiated and developed more than 6,000 years ago.     

<Emphasis Type="SmallCaps">l</Emphasis>-5-hydroxytryptophan resets the circadian locomotor activity rhythm of the nocturnal Indian pygmy field mouse, <Emphasis Type="Italic">Mus terricolor</Emphasis>

We report that l-5-hydroxytryptophan (5-HTP), a serotonin precursor, resets the overt circadian rhythm in the Indian pygmy field mouse, Mus terricolor, in a phase- and dose-dependent manner. We used wheel running to assess phase shifts in the free-running locomotor activity rhythm. Following entrainment to a 12:12 h light–dark cycle, 5-HTP (100 mg/kg in saline) was intraperitoneally administered in complete darkness at circadian time (CT)s 0, 3, 6, 9, 12, 15, 18, and 21, and the ensuing phase shifts in the locomotor activity rhythm were calculated. The results show that 5-HTP differentially shifts the phase of the rhythm, causing phase advances from CT 0 to CT 12 and phase delays from CT 12 to CT 21. Maximum advance phase shift was at CT 6 (1.18 ± 0.37 h) and maximum delay was at CT 18 (−2.36 ± 0.56 h). No extended dead zone is apparent. Vehicle (saline) at any CT did not evoke a significant phase shift. Investigations with different doses (10, 50, 100, and 200 mg/kg) of 5-HTP revealed that the phase resetting effect is dose-dependent. The shape of the phase–response curve (PRC) has a strong similarity to PRCs obtained using some serotonergic agents. There was no significant increase in wheel-running activity after 5-HTP injection, ruling out behavioral arousal-dependent shifts. This suggests that this phase resetting does not completely depend on feedback of the overt rhythmic behavior on the circadian clock. A mechanistic explanation of these shifts is currently lacking.     

Mechanism of Deca-BDE-induced apoptosis in Neuro-2a cells: Role of death-receptor pathway and reactive oxygen species-mediated mitochondrial pathway

Decabromodiphenyl ether (BDE-209) is a prevalent polybrominated diphenyl ether (PBDE) congener known to have neurotoxicity. Effects of BDE-209 on Neuro-2a cells were performed in the present study and the possible apoptotic pathway was discussed. Results indicated that BDE-209 induced Neuro-2a cell apoptosis, increased the protein expression of Fas and Fas-associated death domain-containing protein (FADD) and activated the caspase-8 and -3 activities in a concentration-dependent manner, inferring the death-receptor pathway was involved in the apoptotic process. Meanwhile, BDE-209 exposure increased the Bax/Bcl-2 ratio and decreased the cellular mitochondrial membrane potential (MMP) which led to cytochrome C released to the cytoplasm. The intracellular caspase-9 was elevated simultaneously, which caused downstream caspase cascade and triggered cell apoptosis. Moreover, BDE-209 exposure increased cellular reactive oxygen species (ROS) level in a concentration-dependent manner and the addition of N-acetyl-l-cysteine (NAC), known as ROS scavengers, obviously reduced the apoptotic rate and a positive relationship was observed between the degree of apoptosis blocking and the loss of MMP and ROS production. We thus concluded that BDE-209 induced Neuro-2a cell apoptosis via the combination of the death-receptor signaling pathway and the mitochondrial signaling pathway. The elevated ROS production was considered to magnify the intracellular apoptosis signal and played a crucial role in apoptosis of Neuro-2a cells induced by BDE-209.     

Al nanoparticles: Impact of media and current on the arc fabrication

To optimize the arc fabrication of aluminum nanoparticles (Al Nps), six diverse media were probed including ethylene glycol (EG), distilled water, liquid nitrogen, gaseous nitrogen, sunflower oil, and tap water (all free of any surfactant or stabilizer). These were investigated at three different currents: 50, 100 and 150 A/cm². EG at 50 A/cm² provides the best recipe for the arc fabrication, giving the relatively purest, most dispersed and the smallest size Al Nps (26.6 nm), with the highest yield (91.5 %). Regardless of the medium used, the SEM analyses show that the size increases of Al Nps are directly proportional to the current density employed. The observed sizes for Al Nps in EG (at three different currents/A cm^{- 2}) are 26.6 nm (50), 64.9 nm (100), and 72.5 nm (150). Also, regardless of the current employed, the SEM analyses show different Al Nps size distributions, depending on the medium nature. The XRD extracted average sizes of the arc fabricated Al Nps show reasonable linear correlations with their corresponding elemental weight% or atomic%, acquired through energy-dispersive X-ray spectroscopy (EDX). High quantities of separable industrially significant nanoAl oxides (γ-Al₂O₃, Al(OH)₃) are fabricated along with Al Nps in tap water. In EG medium, TGA revealed only ～7% weight loss, indicating thermal stability of the fabricated Al Nps.     

Effects of different diets on utilization of nitrogen from cattle slurry applied to grassland on a sandy soil in The Netherlands

Dietary adjustments have been suggested as a means to reduce N losses from dairy systems. Differences in fertilizing value of dairy slurry as a result of dietary adjustments were evaluated in a 1-year grassland experiment and by long-term modelling. Slurry composition of non-lactating dairy cows was manipulated by feeding diets with extreme high and low levels of dietary protein and energy. C:N_total ratio of the produced slurries ranged from 5.1 to 11.4. To evaluate their short-term fertilizer N value, the experimental slurries (n = 8) and slurries from commercial farms with variable composition (n = 4), were slit-injected in two grassland fields on the same sandy soil series in the north of The Netherlands (53°10′N, 6°04′E), with differences in sward age and ground water level. The recently established grassland field (NEW) was characterized by lower soil OM, N and moisture contents, less herbs and more modern grass varieties compared to the older grassland field (OLD). Slurry was applied in spring (100 kg N ha⁻¹) and after the first cut (80 kg N ha⁻¹) while in total four cuts were harvested. Artificial fertilizer N treatments were included in the experiment to calculate the mineral fertilizer equivalent (MFE) of slurry N. The OLD field showed a higher total N uptake whereas DM yields were similar for the two fields. Average MFE of the slurries on the OLD field (47%) was lower than on the NEW field (56%), probably as a result of denitrification of slurry N during wet conditions in spring. Slurries from high crude protein diets showed a significantly higher MFE (P < 0.05) compared to low crude protein diets. No significant differences in MFE were observed between slurries from high and low energy diets. On both fields, MFE appeared to be positively related to the ammonium content (P < 0.001) and negatively to the C:N_total ratio of the slurry DM (P = 0.001). Simulation of the effect of long-term annual application of 180 kg N ha⁻¹ with highest and lowest C:N_total ratio suggested that both slurries would lead to an increase in annual soil N mineralization. Both soil N mineralization and SOC appeared to be substantially higher in equilibrium state for the slurry with the highest C:N_total ratio. It is concluded that in a situation with slit-injection, the reduced first-year N availability of slurry with a high C:N_total ratio as observed in the grassland experiment will only be compensated for by soil N mineralization on the very long term.     

区域源解析模式的建立

运用数值分类学手段建立了一种迭代的(iterative)、分裂的(divisive)、不加权的(non-weighting)、非交迭(nonoverlapping)的运算程式(IDNN模式),从实测原始数据中提取可近似作为纯源的样品和相应的性状,通过分析这些近似纯源样品及与之关系密切的其它特征样品的采样参数和成分特征可鉴别或识别源的类型,运用化学质量平衡法(CMB)求解受体样品的定量来源.在对模拟数据的验证处理中,IDNN模式显示了初步成功.