酿酒酵母菌核糖体RNA沉降系数的初步研究

为研究酿酒酵母菌核糖体RNA(rRNA)的沉降系数,用酶解法和液氮研磨法裂解酿酒酵母菌的细胞壁,Trizol Reagent提取其总RNA,同时提取小白鼠和斑马鱼的总RNA进行比较.经紫外分光光度计检测和甲醛琼脂糖变性胶电泳后,RNA纯度好,条带清晰,无弥散或降解现象.试验发现,与酶解法相比,用液氮研磨法破碎酿酒酵母菌细胞壁提取总RNA所用的成本低,时间少,产率和纯度高,适用于少量样品RNA的提取.同时,酿酒酵母菌与斑马鱼和小白鼠总RNA电泳图谱表明,三者的"18S rRNA"在条带大小方面差异较小,而"28S rRNA"差异较大.利用分析型离心机测得的酿酒酵母菌两个较大rRNA的沉降系数分别为24.7S和18.1S.研究结果表明了真核生物rRNA种类的多样性.     

2种调查方法对四川黑竹沟国家级自然保护区3种雉类种群密度调查的比较

种群数量是物种的重要生态学基础资料,合适的密度调查方法是数量估算的基础。2016年4—5月,采用广泛应用于鸡形目Galliformes鸟类种群密度调查的样线法和样点法,调查了四川黑竹沟国家级自然保护区3种鸡形目鸟类(白腹锦鸡Chrysolophus amherstiae、红腹角雉Tragopan temminckii和血雉Ithaginis cruentus)的种群密度。样线法和样点法估算的雄体密度分别是:白腹锦鸡1.20只/km²和(6.31±0.98)只/km²,红腹角雉5.41只/km²和(0.39±0.17)只/km²,血雉3.01只/km²和(5.97±2.70)只/km²。除红腹角雉外,样点法估算的白腹锦鸡、血雉种群密度均大于样线法。建议针对不同鸡形目鸟类采用不同的调查方法,并尽量扩大样本数量,从而提高调查结果的准确性。     

Stability of Prussian Blue in Soils of a Former Manufactured Gas Plant Site

Soil contamination with iron-cyanide complexes is a common problem at former manufactured gas plant (MGP) sites. Dissolution of the cyanide, from Prussian Blue (ferric ferrocyanide), creates an environmental hazard, whereas the risk of groundwater contamination depends on the stability of dissolved iron–cyanide complexes. Lack of a standard leaching method to determine the water-soluble (plant-available) cyanide fraction generates potential limitations for implementing remediation strategies like phytoremediation. Applicability of neutral solution extraction to determine the water-soluble cyanide fraction and the stability of Prussian Blue in surface and near-surface soils of an MGP site in Cottbus, undersaturated and unsaturated water conditions, was studied in column leaching and batch extraction experiments. MGP soils used in the long-term tests varied according to the pH (5.0–7.7) and the total cyanide content (40–1718 mg kg^?1). Column leaching, after four months of percolation, still yielded effluent concentrations exceeding the German drinking water limit (> 50 μg L^?1) and the solubility of Prussian Blue reported in the literature (< 1 mg L^?1) from both alkaline and acidic soils. Long-term (1344 h) extraction of MGP soils with distilled water was sufficient to dissolve 97% of the total cyanide from the slightly alkaline soils and up to 78% from the acidic soils. Both experiments revealed that dissolution of ferric ferrocyanide under circum-neutral pH and oxic water conditions is a function of time, where the released amount is dependent on the soil pH and total cyanide content. Unexpectedly high and continuous solubility of Prussian Blue, both in acidic and slightly alkaline MGP soils, implies the need to introduce an additional cyanide fraction (“readily soluble fraction”) to improve and specify cyanide leaching methods. Long-term extraction of cyanide-contaminated soil in neutral solution seems to be a promising approach to evaluate the potential hazard of groundwater pollution at the MGP sites.     

Comparative study of the antioxidant activity of the essential oils of five plants against the H2O2 induced stress in Saccharomyces cerevisiae

The purpose of this work was to investigate the protective effect of five essential oils (EOs); Rosmarinus officinalis, Thymus vulgaris, Origanum compactum Benth., Eucalyptus globulus Labill. and Ocimum basilicum L.; against oxidative stress induced by hydrogen peroxide in Saccharomyces cerevisiae. The chemical composition of the EOs was analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). The in vitro antioxidant activity was evaluated and the protective effect of EOs was investigated. Yeast cells were pretreated with different concentrations of EOs (6.25–25 µg/ml) for an hour then incubated with H₂O₂ (2 mM) for an additional hour. Cell viability, antioxidants (Catalase, Superoxide dismutase and Glutathione reductase) and metabolic (Succinate dehydrogenase) enzymes, as well as the level of lipid peroxidation (LPO) and protein carbonyl content (PCO) were evaluated. The chemical composition of EOs has shown the difference qualitatively and quantitatively. Indeed, O. compactum mainly contained Carvacrol, O. basilicum was mainly composed of Linalool, T. vulgaris was rich in thymol, R. officinalis had high α-Pinene amount and for E. globulus, eucalyptol was the major compound. The EOs of basil, oregano and thyme were found to possess the highest amount of total phenolic compounds. Moreover, they have shown the best protective effect on yeast cells against oxidative stress induced by H₂O₂. In addition, in a dose dependent manner of EOs in yeast medium, treated cells had lower levels of LPO, lower antioxidant and metabolic enzymes activity than cells exposed to H₂O₂ only. The cell viability was also improved. It seems that the studied EOs are efficient natural antioxidants, which can be exploited to protect against damages and serious diseases related to oxidative stress.     

Confidence intervals for the mean of diagnostic test charge data containing zeros

In this paper, we consider the problem of interval estimation for the mean of diagnostic test charges. Diagnostic test charge data may contain zero values, and the nonzero values can often be modeled by a log-normal distribution. Under such a model, we propose three different interval estimation procedures: a percentile-t bootstrap interval based on sufficient statistics and two likelihood-based confidence intervals. For theoretical properties, we show that the two likelihood-based one-sided confidence intervals are only first-order accurate and that the bootstrap-based one-sided confidence interval is second-order accurate. For two-sided confidence intervals, all three proposed methods are second-order accurate. A simulation study in finite-sample sizes suggests all three proposed intervals outperform a widely used minimum variance unbiased estimator (MVUE)-based interval except for the case of one-sided lower end-point intervals when the skewness is very small. Among the proposed one-sided intervals, the bootstrap interval has the best coverage accuracy. For the two-sided intervals, when the sample size is small, the bootstrap method still yields the best coverage accuracy unless the skewness is very small, in which case the bias-corrected ML method has the best accuracy. When the sample size is large, all three proposed intervals have similar coverage accuracy. Finally, we analyze with the proposed methods one real example assessing diagnostic test charges among older adults with depression.     

粪便标本小肠结肠炎耶尔森菌筛检方法的比较

目的 比较分离培养法、反转录‒聚合酶链反应法对腹泻粪便中小肠结肠炎耶尔森菌的检测情况。方法 在2016年6月至2017年6月临床收治的腹泻患者中选取368例,均使用分离培养法、反转录‒聚合酶链反应法对其粪便中的小肠结肠炎耶尔森菌展开检验,对比分析两种方法的检验结果。结果 反转录‒聚合酶链反应法阳性率为77.17%,比分离培养法的51.09%高,差异有统计学意义（P＜0.05）;反转录‒聚合酶链反应法致病菌株检出率为83.10%,高于分离培养法的59.57%,差异有统计学意义（P＜0.05）。结论 对腹泻患者粪便标本中的小肠结肠炎耶尔森菌检测时,相较于分离培养法,反转录‒聚合酶链反应法阳性检出率更高,可使小肠结肠炎耶尔森菌性腹泻诊断准确率提升。     

Multiple trade‐offs regulate the effects of woody plant removal on biodiversity and ecosystem functions in global rangelands

Woody plant encroachment is a major land management issue. Woody removal often aims to restore the original grassy ecosystem, but few studies have assessed the role of woody removal on ecosystem functions and biodiversity at global scales. We collected data from 140 global studies and evaluated how different woody plant removal methods affected biodiversity (plant and animal diversity) and ecosystem functions (plant production, hydrological function, soil carbon) across global rangelands. Our results indicate that the impact of removal is strongly context dependent, varying with the specific response variable, removal method, and traits of the target species. Over all treatments, woody plant removal increased grass biomass and total groundstorey diversity. Physical and chemical removal methods increased grass biomass and total groundstorey biomass (i.e., non‐woody plants, including grass biomass), but burning reduced animal diversity. The impact of different treatment methods declined with time since removal, particularly for total groundstorey biomass. Removing pyramid‐shaped woody plants increased total groundstorey biomass and hydrological function but reduced total groundstorey diversity. Environmental context (e.g., aridity and soil texture) indirectly controlled the effect of removal on biomass and biodiversity by influencing plant traits such as plant shape, allelopathic, or roots types. Our study demonstrates that a one‐size‐fits‐all approach to woody plant removal is not appropriate, and that consideration of woody plant identity, removal method, and environmental context is critical for optimizing removal outcomes. Applying this knowledge is fundamental for maintaining diverse and functional rangeland ecosystems as we move toward a drier and more variable climate.     

Anoxybacillus sp. SO B1–Immobilized Amberlite XAD-16 for Solid-Phase Preconcentration of Cu(II) and Pb(II) and Their Determinations by Flame Atomic Absorption Spectrometry

A new method for the determination of Cu(II) and Pb(II) by flame atomic absorption spectrometry (FAAS) after preconcentrating on a column containing Anoxybacillus sp. SO B1–immobilized Amberlite XAD-16 was developed. The functional groups of Anoxybacillus sp. SO B1 immobilized on Amberlite XAD-16 were characterized in KBr tablets by Fourier transform infrared (FT-IR) spectrometry. Various parameters such as pH, amount of the adsorbent, eluent type and volume, and flow rate of the sample solution were studied. The optimum pH values of quantitative sorption for Cu(II) and Pb(II) were found to be pH 7.0 and 5.0 and Cu(II) and Pb(II) ions could be quantitatively eluted with 5.0 ml of 1.0 mol L^?1 HCI and 10.0 ml of 0.25 mol L^?1 HNO₃, respectively. Recoveries of Cu(II) and Pb(II) were found to be 100.9 ± 1.57% and 100.3 ± 0.49% (N = 5), the limits of detection of Cu(II) and Pb(II) in the determination by FAAS (3 s, N = 10) were found to be 0.8 and 1.6 μg L^?1, respectively. The proposed enrichment method was applied for metal ion determination from water samples such as two parts of Tigris River water in Diyarbak?r and Elaz??, Lake of Hazar in Elaz??, and tap water in Diyarbak?r. Furthermore, the accuracy of the proposed method was verified by studying the analytical recovery and by analyzing certified reference material (NCS-DC 73350 leaves of poplar).     

Genetic selection for a highly functional cysteine-less membrane protein using site saturation mutagenesis

We describe an efficient method for generating highly functional membrane proteins with variant amino acids at defined positions that couples a modified site saturation strategy with functional genetic selection. We applied this method to the production of a cysteine-less variant of the Crithidia fasciculata inosine-guanosine permease CfNT2 to facilitate biochemical studies using thiol-specific modifying reagents. Of 10 endogenous cysteine residues in CfNT2, two cannot be replaced with serine or alanine without loss of function. High-quality single- and double-mutant libraries were produced by combining a previously reported site saturation mutagenesis scheme based on the Stratagene Quikchange method with a novel gel purification step that effectively eliminated template DNA from the products. Following selection for functional complementation in Saccharomyces cerevisiae cells auxotrophic for purines, several highly functional noncysteine substitutions were efficiently identified at each desired position, allowing the construction of cysteine-less variants of CfNT2 that retained wild-type affinity for inosine. This combination of an improved site saturation mutagenesis technique and positive genetic selection provides a simple and efficient means to identify functional and perhaps unexpected amino acid variants at a desired position.