结核分枝杆菌可视化抗体检测蛋白芯片的制备

目的：利用克隆表达的7种结核分枝杆菌优势表位抗原,建立可视化抗体检测蛋白芯片,用于结核病辅助诊断。方法：将7种结核分枝杆菌优势表位抗原,即38kD、ESAT-6、CFP10、MPT64、Mtb8、Mtb8.4和Mtb16.3点于修饰的基片上,制备可检测7种结核抗体的多靶点蛋白微阵列,建立免疫金银染色检测系统;使用该芯片对48例临床结核病患者血液样品进行检测,并与“金标准”痰涂片（48例）和痰培养（其中的29例）检测结果进行比较,分析其敏感性;对30名献血员血液样品进行检测,分析其特异性。结果：可视化抗体检测蛋白芯片的敏感性分别为98.5％和96.6％,而痰涂片和痰培养检测方法的敏感性分别为35.4％和48.3％;可视化抗体检测蛋白芯片的特异性为93.3％。结论：建立的结核分枝杆菌可视化抗体检测蛋白芯片检测敏感性显著高于痰涂片和痰培养方法,可用于结核病的临床辅助诊断,提高痰涂片和痰培养假阴性的检出率。     

秸秆覆盖免耕土壤真菌群落结构与生态特征研究

不同耕作方法土壤真菌群落结构和生态特征分析表明,翻耕0－10cm土层土壤真菌群落含有29种真菌,其中以Chrysosporium merdarium为优势种;翻耕10－20cm土层含有17种真菌,以Sterile black A为优势种：翻耕20－30cm土层含有10种真菌,其中以Chaetomium bostrychodes为优势种,铁茬0－10cm含16种真菌,其中Sterile black A是优势种;铁茬10－20cm土层含有26种真菌,优势种为Sterile black A;铁茬20－30cm含有14种真菌,其中Chaetomium bostrychodes为优势种,免耕0－10cm土层由23种真菌构成,Trichoderma viride和T.koningii为优势种,免耕10－20cm土层由14个种类构成,Talaromyces trachyspermus为优势种,免耕20－30cm土壤由9种真菌组成,其中Talaromyces trachypermus为优势种,免耕土壤真菌群落的多样性和均匀度指数均较高,主成分分析表明,土壤耕作形成了特征性的真菌区系。     

贵州省后寨河小流域地形因子、人为干扰与石漠化定量研究

为定量研究喀斯特区域地形因子、人为干扰和石漠化之间的关系。基于网格法和实地观测,建立后寨河流域石漠化信息数据库并分析。结果表明:后寨河流域石漠化面积为14.43 km~2,不同利用类型石漠化面积呈以下规律:荒地坡耕地乔木林地耕地灌木林地灌草地草地弃耕地园地乔灌木林地水田经果林地;流域内岩石裸露率和土壤厚度主要受到坡度、坡位和人为干扰的制约,而人为干扰具有主观性,主要在岩石裸露率低、土壤较厚和坡度较缓的区域进行农业生产活动。荒地和弃耕地占据了石漠化面积的32.64%,且存在较大潜在石漠化的风险。回归模型发现人为干扰程度对岩石裸露率(负效应-0.286)和土壤厚度(正效应0.264)的直接效应最大,坡向对岩石裸露率(正效应0.067)和土壤厚度(负效应-0.054)的直接效应最小。认清地形因子、人为干扰和石漠化之间的耦合关系有利于喀斯特小流域石漠化治理。     

马尾松人工林土壤各粒径团聚体湿筛后的有机碳分配

选取25a、45a和65a马尾松人工林为研究对象,采用湿筛法对各粒径土壤团聚体分别湿筛。探究了马尾松人工林各粒径团聚体湿筛后的团聚体有机碳分配,以探讨各粒径团聚体湿筛后分配到同一粒级团聚体有机碳含量及其对团聚体水稳性的贡献差异。结果表明：种植年限增加显著降低土壤团聚体水稳性（P<0.05）;各粒径团聚体湿筛后分配的有机碳随粒级减小含量呈先降后增趋势,以保持原粒级团聚体有机碳（12.96-32.01 g/kg）含量最高,其次是<0.25 mm粒级（8.08-23.53 g/kg）。各粒径团聚体湿筛分配到同一粒级的有机碳以保持原粒级的含量最高（P<0.05）;土壤团聚体水稳性与各粒径团聚体湿筛后保持原粒径的有机碳呈显著或极显著正相关（P<0.05或0.01）,分配到越小的粒级正相关性越不显著。此外,团聚体水稳性与各粒径团聚体湿筛分配到同一粒级的有机碳呈正相关,以保持原粒级相关性最高（P<0.01或0.05）;回归方程及相关性系数表明,有机碳与保持原粒径团聚体呈显著呈或极显著正相关（P<0.05或0.01）,与消散到其他粒级的团聚体呈负相关或极显著负相关（P>0.05或<0.01）。本研究得出有机碳含量增加促进更大粒径团聚体形成。反之,促使大粒径团聚体向较小粒径团聚体转化。同一粒级团聚体间,保持原粒级团聚体比易转化形成更大粒级团聚体有更高的有机碳含量和更强的水稳性,这对团聚体的固碳提供了新的认识。     

Constraining oceanic oxygenation during the Shuram excursion in South China using thallium isotopes

Ediacaran sediments record an unusual global carbon cycle perturbation that has been linked to widespread oceanic oxygenation, the Shuram negative C isotope excursion (NCIE). However, proxy‐based estimates of global ocean redox conditions during this event have been limited largely due to proxy specificity (e.g., euxinic sediments for Mo and U isotopes). Modern global seawater documents a homogenous Tl isotope composition (ε²⁰⁵Tl = ?6.0) due to significant manganese oxide burial, which is recorded in modern euxinic sediments. Here, we provide new data documenting that sediments deposited beneath reducing but a non‐sulfidic water column from the Santa Barbara Basin (ε²⁰⁵Tl = ?5.6 ± 0.1) also faithfully capture global seawater Tl isotope values. Thus, the proxy utilization of Tl isotopes can extend beyond strictly euxinic settings. Second, to better constrain the global redox conditions during the Shuram NCIE, we measured Tl isotopes of locally euxinic and ferruginous shales of the upper Doushantuo Formation, South China. The ε²⁰⁵Tl values of these shales exhibit a decreasing trend from ≈?3 to ≈?8, broadly coinciding with the onset of Shuram NCIE. There are ε²⁰⁵Tl values (?5.1 to ?7.8) during the main Shuram NCIE interval that approach values more negative than modern global seawater. These results suggest that manganese oxide burial was near or even greater than modern burial fluxes, which is likely linked to an expansion of oxic conditions. This ocean oxygenation may have been an important trigger for the Shuram NCIE and evolution of Ediacaran‐type biota. Subsequently, Tl isotopes show an increasing trend from the modern ocean value to values near the modern global inputs or even heavier (ε²⁰⁵Tl ≈ ?2.5 ~ 0.4), occurring prior to recovery from the NCIE. These records may suggest that there was a decrease in the extent of oxygenated conditions in the global oceans during the late stage of the Shuram NCIE.     

UHRF2 commissions the completion of DNA demethylation through allosteric activation by 5hmC and K33-linked ubiquitination of XRCC1

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Production of recombinant Bombyx mori nucleopolyhedrovirus in silkworm by intrahaemocoelic injection with invasive diaminopimelate auxotrophic Escherichia coli containing BmNPV-Bacmid

The present study elaborates a cost-effective and transfectant-free method for generating recombinant Bombyx mori (silkworm) nucleopolyhedrovirus in silkworm larvae and pupae by injecting invasive Escherichia coli carrying BmBacmid [BmNPV (B. mori nucleopolyhedrovirus)-Bacmid] into larval haemocoel. Up to 109 PFU (plaque-forming units)/ml of infective recombinant baculovirus was generated in the silkworm by intrahaemocoelic injection with 106 DAP (diaminopimelic acid) auxotrophic and BmBacmid containing E. coli cells expressing both invasin and listeriolysin. Thus 1?ml of overnight culture of E. coli is sufficient to inject more than 2000 larvae, while DAP costing up to $1 is enough to inject about 4000 larvae. Recombinant proteins can be controlled to be expressed mainly in pupae by adjusting the injection dose, too. In this new method, many original manipulations have been eliminated, including BmBacmid preparation and the subsequent complex transfection procedures. Hence it is a time- and cost-saving means for large-scale injection of B. mori for recombinant baculovirus production in comparison with the traditional transfection methods, which may play an important role in the industrial development of the BmNPV-silkworm bioreactor.     

Species delineation using Bayesian model-based assignment tests: a case study using Chinese toad-headed agamas (genus <Emphasis Type="Italic">Phrynocephalus</Emphasis>)

Background  

Species are fundamental units in biology, yet much debate exists surrounding how we should delineate species in nature. Species discovery now requires the use of separate, corroborating datasets to quantify independently evolving lineages and test species criteria. However, the complexity of the speciation process has ushered in a need to infuse studies with new tools capable of aiding in species delineation. We suggest that model-based assignment tests are one such tool. This method circumvents constraints with traditional population genetic analyses and provides a novel means of describing cryptic and complex diversity in natural systems. Using toad-headed agamas of the Phrynocephalus vlangalii complex as a case study, we apply model-based assignment tests to microsatellite DNA data to test whether P. putjatia, a controversial species that closely resembles P. vlangalii morphologically, represents a valid species. Mitochondrial DNA and geographic data are also included to corroborate the assignment test results.     

Akita Spontaneously Type 1 Diabetic Mice Exhibit Elevated Vascular Arginase and Impaired Vascular Endothelial and Nitrergic Function

Background

Elevated arginase (Arg) activity is reported to be involved in diabetes-induced vascular endothelial dysfunction. It can reduce L-arginine availability to nitric oxide (NO) synthase (NOS) and NO production. Akita mice, a genetic non-obese type 1 diabetes model, recapitulate human diabetes. We determined the role of Arg in a time-course of diabetes-associated endothelial dysfunction in aorta and corpora cavernosa (CC) from Akita mice.

Methods and Results

Endothelium-dependent relaxation, Arg and NOS activity, and protein expression levels of Arg and constitutive NOS were assessed in aortas and CC from Akita and non-diabetic wild type (WT) mice at 4, 12 and 24 wks of age. Systolic blood pressure (SBP) was assessed by tail cuff. In aorta and CC, Akita mice exhibited a progressive impairment of vascular endothelial and nitrergic function increased Arg activity and expression (Arg1 in aorta and both Arg1 and Arg2 in CC) compared with that of age-matched WT mice. Treatment of aorta and CC from Akita mice with an Arg inhibitor (BEC or ABH) reduced diabetes-induced elevation of Arg activity and restored endothelial and nitrergic function. Reduced levels of phospho-eNOS at Ser¹¹⁷⁷ (in aorta and CC) and nNOS expression (in CC) were observed in Akita mice at 12 and 24 wks. Akita mice also had decreased NOS activity in aorta and CC at 12 and 24 wks that was restored by BEC treatment. Further, Akita mice exhibited moderately increased SBP at 24 wks and increased sensitivity to PE-induced contractions in aorta and sympathetic nerve stimulation in CC at 12 and 24 wks.

Conclusions

Over 24 wks of diabetes in Akita mice, both aortic and cavernosal tissues exhibited increased Arg activity/expression, contributing to impaired endothelial and nitrergic function and reduced NO production. Our findings demonstrate involvement of Arg activity in diabetes-induced impairment of vascular function in Akita mouse.