Some Molecular and Cellular Stress Mechanisms Associated with Neurodegenerative Diseases and Atherosclerosis

Chronic stress is a combination of nonspecific adaptive reactions of the body to the influence of various adverse stress factors which disrupt its homeostasis, and it is also a corresponding state of the organism’s nervous system (or the body in general). We hypothesized that chronic stress may be one of the causes occurence of several molecular and cellular types of stress. We analyzed literary sources and considered most of these types of stress in our review article. We examined genes and mutations of nuclear and mitochondrial genomes and also molecular variants which lead to various types of stress. The end result of chronic stress can be metabolic disturbance in humans and animals, leading to accumulation of reactive oxygen species (ROS), oxidative stress, energy deficiency in cells (due to a decrease in ATP synthesis) and mitochondrial dysfunction. These changes can last for the lifetime and lead to severe pathologies, including neurodegenerative diseases and atherosclerosis. The analysis of literature allowed us to conclude that under the influence of chronic stress, metabolism in the human body can be disrupted, mutations of the mitochondrial and nuclear genome and dysfunction of cells and their compartments can occur. As a result of these processes, oxidative, genotoxic, and cellular stress can occur. Therefore, chronic stress can be one of the causes forthe occurrence and development of neurodegenerative diseases and atherosclerosis. In particular, chronic stress can play a large role in the occurrence and development of oxidative, genotoxic, and cellular types of stress.     

Influencer-defaulter mutation-based optimization algorithms for predicting electricity prices

Efficient electricity price forecasting plays a significant role in our society. In this paper, a novel influencer-defaulter mutation (IDM) mutation operator has been proposed. The IDM operator has been combined with six well-known optimization algorithms to create mutated optimization algorithms whose performance has been tested on twenty-four standard benchmark functions. Further, the artificial neural network is integrated with mutated optimization algorithms to solve the electricity price prediction problem. The policymakers can identify appropriate variables based on the predicted prices to help future market planning. The statistical results prove the efficacy of the IDM operator on the recent optimization algorithms.     

New Insights into the Pathogenesis of Systemic Mastocytosis

Mastocytosis is a type of myeloid neoplasm characterized by the clonal, neoplastic proliferation of morphologically and immunophenotypically abnormal mast cells that infiltrate one or more organ systems. Systemic mastocytosis (SM) is a more aggressive variant of mastocytosis with extracutaneous involvement, which might be associated with multi-organ dysfunction or failure and shortened survival. Over 80% of patients with SM carry the KIT D816V mutation. However, the KIT D816V mutation serves as a weak oncogene and appears to be a late event in the pathogenesis of mastocytosis. The management of SM is highly individualized and was largely palliative for patients without a targeted form of therapy in past decades. Targeted therapy with midostaurin, a multiple kinase inhibitor that inhibits KIT, has demonstrated efficacy in patients with advanced SM. This led to the recent approval of midostaurin by the United States Food and Drug Administration and European Medicines Agency. However, the overall survival of patients treated with midostaurin remains unsatisfactory. The identification of genetic and epigenetic alterations and understanding their interactions and the molecular mechanisms involved in mastocytosis is necessary to develop rationally targeted therapeutic strategies. This review briefly summarizes recent developments in the understanding of SM pathogenesis and potential treatment strategies for patients with SM.     

Dihydrofolate Reductase Thermosensitive (ts-dfrA) Mutant Induces Dihydrofolate Overproduction by Lactobacillus plantarum

Dihydrofolate (dihydrofolic acid, vitamin B9) is one of the principle reduced forms of folates, which is converted to tetrahydrofolate by dihydrofolate reductase (DHFR). The aim of the present study was to overproduce total folate by inducing accumulation of dihydrofolate (DHF) due to inhibition of DHFR in a thermosensitive mutant (TS) of Lactobacillus plantarum. This mutant (TS34) was obtained by UV-mutagenesis of L. plantarum wild-type parent strain (HMA07) isolated from fermented sausage. The mutant TS34 was characterized by its ability to grow at 30°C but not at 40°C. Strain HMA07 and its mutant TS34 were identified by determining partial sequence of 16S rRNA gene. A single C→T exchange in the dfrA (dihydrofolate reductase gene) at nucleotide 184 was detected. This exchange led to the conversion of leucine (L) to phenylalanine (F) at the position 62 in amino acid sequence (L62F transition). TS43 showed significant (p < 0.05) reduction in DHFR activity from 1.51 to 0.06 Unit/mg protein, resulting in increasing DHF amount at the expense of THF. Accordingly, total folate production by the mutant TS34 (in two-stage fermentation process with temperature shift-up from 30°C to 40°C) increased by three-fold compared with the parental strain HMA07. In conclusion, thermosensitive mutation in dfrA caused inhibition of DHFR and resulted in accumulation of DHF. The technique described here could be considered as novel strategy for overproduction of DHF in two-stage fermentation process.     

Synthesis,structure, CO oxidation,and H2 production activities of CaCu3−xMnxTi4−xMnxO12 (x = 0, 0.5, and 1.0)

Synthesis of nanocrystalline pristine and Mn-doped calcium copper titanate quadruple perovskites, CaCu_3?xMn_xTi_4?xMn_xO₁₂ (x = 0, 0.5, and 1.0) by modified citrate solution combustion method has been reported. Powder X-ray diffraction patterns attest the phase purity of the perovskite materials. Average particle sizes of all the materials obtained from the Scherrer's formula are in the range of 55–70 nm. The specific surface areas for all the perovskites obtained from BET isotherms are found to be low as expected for the condensed oxide systems and fall in the range of 13–17 m² g^?1. Transmission electron microscopy studies show a reduction in particle size of CaCu₃Ti₄O₁₂ with increase in Mn doping. Ca and Ti are present in +2 and +4 oxidation states in all the materials as demonstrated by X-ray photoelectron spectroscopy analyses. Cu²⁺ gets reduced in CaCu₃Ti₄O₁₂ with higher Mn content. Mn is observed to be present only in +3 oxidation state. All the materials have been examined to be active in CO oxidation as well as H₂ production from methanol steam reforming. CaCu₃Ti₄O₁₂ with ~14 at.% Mn is found to show best catalytic activities among these materials. A comprehensive analysis of the catalytic activities of these perovskites toward CO oxidation and H₂ production from MSR reveal the cooperative activity of copper-manganese in the doped perovskites and it is more effective at lower manganese content.     

碳源对manA基因突变大肠杆菌代谢的影响

ManA基因编码的甘露糖-6-磷酸异构酶在大肠杆菌中催化D-甘露糖和D-果糖的异构化,促进大肠杆菌对碳源的代谢吸收。本文通过研究manA基因突变大肠杆菌对碳源的利用和编码糖代谢基因情况,探讨甘露糖-6-磷酸异构酶对大肠杆菌糖代谢的影响。采用Ⅱ型内含子逆转录突变方法构建manA基因突变大肠杆菌,分析manA基因突变大肠杆菌对不同碳源的利用情况和manA基因突变对大肠杆菌糖代谢相关基因表达的影响,结果显示,大肠杆菌BL21(DE3)ΔmanA以甘露糖、果糖为碳源时,菌株生长受到显著抑制;以淀粉为碳源时,BL21(DE3)ΔmanA菌株的生长显著优于野生型大肠杆菌;以葡萄糖为碳源时,manA基因突变对大肠杆菌的生长无显著影响。通过基因表达分析,发现大肠杆菌BL21(DE3)ΔmanA中甘露糖代谢相关基因的表达显著性降低;果糖代谢途径中6-磷酸果糖激酶Ⅰ亚基的编码基因(pfkA)显著下调表达;水解淀粉的α-淀粉酶编码基因(malS)显著性上调表达。ManA基因突变影响大肠杆菌甘露糖、果糖和淀粉代谢途径中相关基因的表达,从而影响大肠杆菌对碳源的利用。     

常压室温等离子体(ARTP)诱变选育高核酸酿酒酵母

核糖核酸(RNA)是一类非常重要的生物分子,降解后得到的核苷酸、核苷及碱基具有广泛用途。酿酒酵母是目前生产RNA的主要食品级微生物。本研究采用常压室温等离子体(ARTP)技术进行酿酒酵母诱变育种,利用氯化钾敏感性筛选,多次反复诱变最终得到在以糖蜜为碳源的摇瓶试验中RNA含量为112 mg-RNA/g-DCW,提高了39%的突变菌株Y17aM3。经过对Y17aM3培养条件优化后,确定生产RNA最适接种量为10%,最适p H为5.5,最适温度为26℃,且传代稳定性良好。研究发现在最佳培养条件下,添加磷酸可使Y17aM3的RNA含量提高至119 mg-RNA/g-DCW;添加蛋白胨可使Y17aM3的RNA含量提高至122 mg-RNA/g-DCW。上述结果不仅证明ARTP诱变育种方法突变效果显著,可应用于工业微生物的选育,而且有助于降低基于核苷酸的食品添加剂的生产成本。     

瓦斯抽采钻孔水射流协同修护技术研究与应用

瓦斯抽采钻孔普遍存在因变形、煤渣积聚及塌孔等导致钻孔堵塞和抽采效果差的问题。通过分析钻孔塌堵失稳机制,得出煤岩体性质、地质构造及多应力耦合条件是造成钻孔失稳的主要因素,进而推断相应堵孔段情形。利用高压水射流解堵作用,提出了水射流疏通-筛管护孔协同修护技术,并研制出轻型气动钻孔修复装备。应用结果表明,该协同修护技术能有效解决瓦斯抽采钻孔塌堵后无法有效抽采的技术难题,试验钻孔修护深度达到50 m,修护完成后单孔抽采瓦斯浓度和瓦斯纯流量比修复前提高0.57~3.67倍和0.99~5.15倍,抽采效果大幅改善,实现了塌堵钻孔的快速便捷修复进而确保了瓦斯流动通道的畅通。     

盖子环替换及定点突变提高菜豆环氧化物水解酶对邻甲基苯基缩水甘油醚的催化性能

菜豆环氧化物水解酶1和2（PvEH1、PvEH2）能够动力学拆分外消旋邻甲基苯基缩水甘油醚（rac-oMGE），从而保留(R)-oMGE。基于对PvEH1和PvEH2结构的同源模拟和分析，发现二者分子中的盖子环差异较大，故本文选择盖子环作为研究目标。经融合聚合酶链式反应（FPCR），获得了PvEH2的盖子环区域被PvEH1对应区域替换的杂合酶Pv2Pv1。用全细胞酶E. coli/pv2pv1催化rac-oMGE，当(S)-oMGE刚好水解完全时，产物(S)-3-邻甲苯基-1,2-丙二醇（(S)-oTPD）的ee_p由PvEH2的58.3%提高至75.5%。为进一步提高酶的性质，在Pv2Pv1中选取11个氨基酸位点进行丙氨酸(A)突变，获得最优突变子E. coli/pv2pv1^K176A，活性为E. coli/pv2pv1（4.2U/g）的2.1倍，且当S构型的底物刚好完全水解时，(S)-oTPD的ee_p进一步提高为80.3%。分子对接分析发现，盖子环替换和K176位点突变为A，均使(R)-oMGE环氧环中的C_α更易受到酶中D101位点的攻击。利用E. coli/pv2pv1^K176A催化150mmol/L rac-oMGE水解制备(R)-oMGE（ee_s＞99%）和(S)-oTPD（ee_p=80.4%），二者的产率Y_S和Y_P分别为32.7%和60.1%，时空产率STY_S和STY_P为1.6g/(L·h)和3.3g/(L·h)。本实验为改善EH的催化性质提供了一种有效策略。     

一种大型复杂构件加工新模式及新装备探讨

大型复杂构件是航空航天、能源、船舶等领域装备的核心结构件，此类构件通常具有尺寸大、形状复杂、刚性弱等特点。传统“分体离线加工-在线检测”模式存在工艺不稳定、过程复杂、柔性差、周期长等问题，以龙门式多轴数控机床加工为代表的“包容式”加工模式，难以适应大型复杂构件的高效高质量加工制造需求。提出一种基于移动式和吸附式机器人的多机协同原位加工新模式，通过多机器人系统自主寻位、精确定位加工与加工质量原位检测，实现大型复杂构件多安装面并行铣削、制孔与打磨等作业。多机器人系统包括移动式混联机器人、吸附式并联机器人、移动式串联铣削机器人、移动式双臂加工机器人和移动式打磨机器人。构建多机协同原位加工模式，需要揭示多机器人协同原位加工行为与大型弱刚性结构件质量控制的交互机理，面临着本体、测量、工艺和集成四个方面的挑战，需要设计高灵活、高刚度的移动式和吸附式加工机器人，解决移动机器人自主准确寻位和超大结构件原位高精检测难题，攻克加工变形误差在线补偿和振动抑制技术，通过集成实现多机协同高效高精加工，为大型复杂构件的高效高质量制造提供创新技术及装备，并实现此类构件制造核心技术及装备自主可控。