Genome-Based Metabolic Engineering of Mannheimia succiniciproducens for Succinic Acid Production

Succinic acid is a four-carbon dicarboxylic acid produced as one of the fermentation products of anaerobic metabolism. Based on the complete genome sequence of a capnophilic succinic acid-producing rumen bacterium, Mannheimia succiniciproducens, gene knockout studies were carried out to understand its anaerobic fermentative metabolism and consequently to develop a metabolically engineered strain capable of producing succinic acid without by-product formation. Among three different CO₂-fixing metabolic reactions catalyzed by phosphoenolpyruvate (PEP) carboxykinase, PEP carboxylase, and malic enzyme, PEP carboxykinase was the most important for the anaerobic growth of M. succiniciproducens and succinic acid production. Oxaloacetate formed by carboxylation of PEP was found to be converted to succinic acid by three sequential reactions catalyzed by malate dehydrogenase, fumarase, and fumarate reductase. Major metabolic pathways leading to by-product formation were successfully removed by disrupting the ldhA, pflB, pta, and ackA genes. This metabolically engineered LPK7 strain was able to produce 13.4 g/liter of succinic acid from 20 g/liter glucose with little or no formation of acetic, formic, and lactic acids, resulting in a succinic acid yield of 0.97 mol succinic acid per mol glucose. Fed-batch culture of M. succiniciproducens LPK7 with intermittent glucose feeding allowed the production of 52.4 g/liter of succinic acid, with a succinic acid yield of 1.16 mol succinic acid per mol glucose and a succinic acid productivity of 1.8 g/liter/h, which should be useful for industrial production of succinic acid.     

Genetic and phylogenetic analyses of human T-cell lymphotropic virus type I variants from Melanesians with an without spastic myelopathy

Molecular variants of human T-cell lymphotropic virus type I (HTLV-I) have been isolated recently from lifelong residents of remote Melanesian populations, including a Solomon Islander with tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM) or HTLV-I myeloneuropathy. To clarify the genetic heterogeneity and molecular epidemiology of disease-associated strains of HTLV-I, we enzymatically amplified, then directly sequenced representative regions of thegag, pol, env, andpX genes of HTLV-I strains from Melanesians with and without TSP/HAM, and aligned and compared these sequences with those of HTLV-I strains from patients with TSP/HAM or adult T-cell leukemia/lymphoma and from asymptomatic carriers from widely separated and culturally disparate populations. Overall, the HTLV-I variant from the Solomon Islander with TSP/HAM, like HTLV-I strains from asymptomatically infected Melanesians, diverged by approx 7% from cosmopolitan HTLV-I strain. No disease-specific viral sequences were found. Gene phylogenies, as determined by the unweighted pair-goup method of assortment and by the maximum parsimony method, indicated that the Melanesian and cosmopolitan strains of HTLV-I have evolved along separate geographically dependent lineages, one comprised of HTLV-I strains from Papua New Guinea and the Solomon Islands, and the other composed of virus strains from Japan, India, the Caribbean, Polynesia, the Americas, and Africa. The total absence of nonhuman primates in Papua New Guinea and the Solomon Islands precludes any possibility that the Melanesian HTLV-I strains have evolved recently from the simian homolog of HTLV-I.     

Identification and characterization of antifungal active substances of Streptomyces hygroscopicus BS-112

An antifungal Actinomyces BS-112 strain, with Aspergillus flavus as the target pathogen, was isolated from soil in the forest land of Mountain Tai. This strain showed a strong antagonistic activity against various mold fungi in food and feed. Strain BS-112 was identified as Streptomyces hygroscopicus based on its morphologic, cultural, physiological, biochemical characteristics, cell wall components and 16S rDNA sequence. Four active components were separated and purified from strain BS-112. These four antifungal components were identified as tetrins A and B and tetramycins A and B using spectroscopic analysis including mass spectrometry and nuclear magnetic resonance spectroscopy. Tetrins A and B and tetramycins A and B strongly inhibited the growth of A. flavus, A. alutaceus, A. niger, and A. fumigatus in vitro.     

功能区胶质瘤患者初级运动皮层激活的偏侧化及运动网络功能连接研究

无运动障碍的功能区胶质瘤患者,其手术风险较高,结合手运动任务态功能磁共振(fMRI)及运动网络功能连接能否更准确的评估手术风险需要进一步探索.本研究收集24例运动功能区胶质瘤且无明显肢体运动功能障碍的患者,非运动功能区胶质瘤患者8例,术前进行fMRI手运动任务态及静息态检查,术后3个月对患者进行随访.计算初级运动皮质(M1)激活的偏侧化指数(lateralization indices,LI).选取运动网络感兴趣区域(ROI),计算双侧M1与各ROI间功能连接系数(FC).运用受试者工作特性曲线(receiver operating characteristic curve,ROC)评估M1激活的LI对术后偏瘫的预测效能.同非运动功能区胶质瘤患者相比,运动功能区胶质瘤患者M1激活LI显著升高(P=0.001).同术后未偏瘫功能区胶质瘤患者相比,术后偏瘫患者M1激活LI值显著升高(P=0.011).ROC曲线下面积(AUC)=0.867,临界点LI=0.31,LI≥0.31对术后偏瘫预测灵敏性为87.5%,特异性为87.5%.以M1激活LI≥0.31将病人分为手术高风险组及低风险组,手术高风险组患者双侧M1与运动网络多个ROI间FC出现显著改变.本研究中功能区胶质瘤患者虽无肢体运动功能障碍,但肿瘤对功能区皮层激活及运动网络FC已有不同程度的破坏;结合任务态及静息态fMRI可以更好地评估手术风险并了解机体功能受损及代偿机制.     

Determination of 3-keto-4-ene steroids and their hydroxylated metabolites catalyzed by recombinant human cytochrome P450 1B1 enzyme using gas chromatography-mass spectrometry with trimethylsilyl derivatization

A protocol utilizing gas chromatography with selected ion monitoring mass spectrometric detection (GC-SIM-MS) using a simplified trimethylsilyl (TMS) derivatization protocol was developed and validated for the determination of hydroxylated metabolites of 3-keto-4-ene steroids such as testosterone, progesterone and androstenedione. Hydroxylated metabolites catalyzed by human CYP1B1 were extracted with methylene chloride and derivatized with BSTFA-10% TMCS. To get an optimal derivatizing condition, the effect of various incubation times and temperatures was evaluated. When the incubation temperature and time in the presence of the TMS derivatizing agent were increased, the 3-keto group became derivatized with TMS to form a 3-TMS derivative. To minimize the formation of the TMS ether on the 3-keto group, a reaction condition of 56 degrees C for 10 min was used for the routine measurement of the steroids and their hydroxylated metabolite. Performance studies including linearity of calibration curves, extraction efficiency and precision were performed. Linearity of the calibration curves was satisfactory from 0.125 to 5 microM for most compounds except 21-hydroxyprogesterone and 16alpha-hydroxyandrostenedione which deviated from linearity at the lower concentrations. Mean percentage extraction recoveries were greater than 80% for all compounds. Most compounds showed good precisions with C.V.s of within-day precision of less than 5% and C.V.s of between-day precision of less than 10%. The selected ion chromatograms from the recombinant human CYP1B1 incubations with testosterone, progesterone and androstenedione showed evidence of 6beta-, 16alpha-, 2alpha-, and 15alpha-hydroxytestosterone, 6alpha- and 16alpha-hydroxyprogesterone and 6alpha- and 16alpha-hydroxyandrostenedione, respectively. There was no significant interference associated with Escherichia coli membrane extracts in detecting hydroxylated metabolites. This procedure provides a rapid and sensitive method for the evaluation of steroid hydroxylation by CYP isoenzymes.     

海南俄贤岭紫毛野牡丹灌丛群落特征研究

采用样方调查方法,对海南俄贤岭紫毛野牡丹（Melastoma penicillatum Naud.）灌丛群落的种类组成、区系类型和群落结构等进行了研究。结果显示,在200 m²的样地内,有维管植物37种,隶属于26科33属;属的区系成分以热带亚洲（印度-马来西亚）分布和泛热带分布为主,分别占非世界总属数的31.03%和24.14%。在种的水平上,泛热带分布最多,占非世界分布总种数的25%。群落种类主要由灌木（24种）和草本（9种）构成,灌木层中以紫毛野牡丹为优势种,草本层以乌毛蕨（Blechnum orientale L.）为优势种;藤本植物仅有3种。生活型以中高位芽为主,占40.54%;群落的频度划分为5个等级（A：1%~20%、B：21%~40%、C：41%~60%、D：61%~80%、E：81%~100%）,频度等级的分布规律为A > B > D > C=E。与该地海南凤仙花（Impatiens hainanensis Y.L.Chen）群落和海南大戟（Euphorbia hainanensis Croizat）灌丛群落进行比较发现,该群落植物种类更少,分布更稀疏,层次更明显。     

A Fully Verified Theoretical Analysis of Contact‐Mode Triboelectric Nanogenerators as a Wearable Power Source

Harvesting mechanical energy from human activities by triboelectric nanogenerators (TENGs) is an effective approach for sustainable, maintenance‐free, and green power source for wireless, portable, and wearable electronics. A theoretical model for contact‐mode triboelectric nanogenerators based on the principles of charge conservation and zero loop‐voltage is illustrated. Explicit expressions for the output current, voltage, and power are presented for the TENGs with an external load of resistance. Experimental verification is conducted by using a laboratory‐fabricated contact‐mode TENG made from conducting fabric electrodes and polydimethylsiloxane/graphene oxide composite as the dielectric layer. Excellent agreements of the output voltage, current, and power are demonstrated between the theoretical and experimental results, without any adjustable parameters. The effects of the moving speed on output voltage, current, and power are illustrated in three cases, that is, the motion with constant speed, the sinusoidal motion cycles, and the real walking cycles by human subject. The fully verified theoretical model is a very powerful tool to guide the design of the device structure and selection of materials, and optimization of performance with respect to the application conditions of TENGs.     

越冬期测墒补灌对小麦耗水特性和光合有效辐射截获利用的影响

于2013—2014和2014—2015年两个小麦生长季进行田间试验,供试品种为‘济麦22’,设置5个处理,分别为W₀(全生育期不灌水)、W₁(越冬期不灌水,拔节期和开花期分别补灌至0～40 cm土层土壤相对含水量为65%和70%)、W₂(越冬期、拔节期和开花期分别补灌至土壤相对含水量为70%、65%和70%)、W₃(越冬期、拔节期和开花期分别补灌至土壤相对含水量为75%、65%和70%)和W₄(越冬期、拔节期和开花期均定量灌溉60 mm),研究越冬期测墒补灌对小麦耗水特性和光合有效辐射截获利用的影响.结果表明: 总灌水量及其占总耗水量的比例为W₄＞W₃＞W₂＞W₁＞W₀;土壤贮水消耗量占总耗水量的比例为W₀＞W₁、W₂＞W₃、W₄;总耗水量和开花至成熟期的耗水量均为W₄＞W₂、W₃＞W₁＞W₀.两生长季小麦开花后冠层光合有效辐射(PAR)截获率为W₄＞W₂、W₃＞W₁＞W₀,而花后冠层PAR反射率各处理间的表现与之相反.灌水处理中干物质净积累量为W₄处理最高,W₁处理最低.两生长季小麦越冬期0～40 cm土层土壤相对含水量补灌至70%的W₂处理籽粒产量仅低于定量灌溉的W₄处理,水分利用效率和灌溉效益最高,是本试验条件下节水高产的最优处理.     

Colonic epithelium rejuvenation through YAP/TAZ

Stem cells are critical for normal tissue homeostasis and injury‐induced regeneration, but how wounding affects tissue regeneration at the cellular and molecular levels is not yet fully understood. A new study by Yui et al ( 2018 ) demonstrates that the extracellular matrix (ECM) remodeling modulates intestinal stem cells in tissue repair and regeneration via activation of the Hippo pathway effectors YAP/TAZ.