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71.
Many N2-fixing organisms can turn off nitrogenase activity in the presence of NH4
+ and turn it on again when the NH4
+ is exhausted. One of the most interesting systems for accomplishing this is by covalent modification of one subunit of dinitrogenase reductase by dinitrogenase reductase ADP-ribosyltransferase (DRAT). The system can be reactivated when NH4
+ is exhausted, by dinitrogenase reductase activating glycohydrolase (DRAG) which removes the inactivating group. It is fascinating that some species of the genusAzospirillum possess the DRAT and DRAG systems (A. lipoferum andA. brasilense), whereasA. amazonense in the same genus lacks DRAT and DRAG.A. amazonense responds to NH4
+ but does not exhibit modification of dinitrogenase reductase characteristic of the action of DRAT. However, it has been possible to clone DRAT and DRAG and to introduce them intoA. amazonense, whereupon they become functional in this organism. The DRAT and DRAG system does not appear to function inAcetobacter diazotrophicus, an organism isolated from sugar cane, that fixes N2 at a pH as low as 3.0.A. diazotrophicus does show a rather sluggish response to NH4
+. A level of about 10 M NH4
+ is required to switch off the system. The response to NH4
+ is influenced by the dissolved oxygen concentration (DOC) as has been reported forAzospirillum sp. A DOC in equilibrium with 0.1 to 0.2 kPa O2 seems optimal for the response inA. diazotrophicus. 相似文献
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74.
The use of voluntary assisted dying as an end‐of‐life option has stimulated concerns and debates over the past decades. Although public attitudes towards voluntary assisted dying (including euthanasia and physician‐assisted suicide) are well researched, there has been relatively little study of the different reasons, normative reasoning and rhetorical strategies that people invoke in supporting or contesting voluntary assisted dying in everyday life. Using a mix of computational textual mining techniques, keyword study and qualitative thematic coding to analyse public submissions to a parliamentary inquiry into voluntary assisted dying in Australia, this study critically examines the different reasons, normative reasoning and rhetorical strategies that people invoke in supporting or contesting voluntary assisted dying in everyday life. The analysis identified complex and potentially contradictory ethical principles being invoked on both sides of the debate. These findings deepen our understanding of the moral basis of public reasoning about end‐of‐life matters and will help to inform future discussions on policy and law reform. The findings underscore the importance of sound normative reasoning and the use of caution when interpreting opinion polls to inform policy. 相似文献
75.
Ayal Schaffer Mark Sinyor Paul Kurdyak Simone Vigod Jitender Sareen Catherine Reis Diane Green James Bolton Anne Rhodes Sophie Grigoriadis John Cairney Amy Cheung 《World psychiatry》2016,15(2):135-145
The objective of this study was to detail the nature and correlates of mental health and non‐mental health care contacts prior to suicide death. We conducted a systematic extraction of data from records at the Office of the Chief Coroner of Ontario of each person who died by suicide in the city of Toronto from 1998 to 2011. Data on 2,835 suicide deaths were linked with provincial health administrative data to identify health care contacts during the 12 months prior to suicide. Sub‐populations of suicide decedents based on the presence and type of mental health care contact were described and compared across socio‐demographic, clinical and suicide‐specific variables. Time periods from last mental health contact to date of death were calculated and a Cox proportional hazards model examined covariates. Among suicide decedents, 91.7% had some type of past‐year health care contact prior to death, 66.4% had a mental health care contact, and 25.3% had only non‐mental health contacts. The most common type of mental health contact was an outpatient primary care visit (54.0%), followed by an outpatient psychiatric visit (39.8%), an emergency department visit (31.1%), and a psychiatric hospitalization (21.0%). The median time from last mental health contact to death was 18 days (interquartile range 5‐63). Mental health contact was significantly associated with female gender, age 25‐64, absence of a psychosocial stressor, diagnosis of schizophrenia or bipolar disorder, past suicide attempt, self‐poisoning method and absence of a suicide note. Significant differences between sub‐populations of suicide decedents based on the presence and nature of their health care contacts suggest the need for targeting of community and clinical‐based suicide prevention strategies. The predominance of ambulatory mental health care contacts, often close to the time of death, reinforce the importance of concentrating efforts on embedding risk assessment and care pathways into all routine primary and specialty clinical care, and not only acute care settings. 相似文献
76.
对生物体内已有的或者人工组装的生物合成途径进行优化操作涉及两个重要问题:代谢途径中关键酶的活性及蛋白表达水平。对于酶表达水平的研究,传统的做法是采用强启动子控制下的靶蛋白过量表达策略。靶蛋白的过量表达通常会导致细胞内积累大量的无活性包涵体,从而严重影响细胞的生理状态和相关生物途径的有效运转。针对这一问题,设计一种分子开关来精确调控生物合成过程中关键酶的表达水平,对于研究生物合成途径的代谢节律以及促进生物合成途径高效运转都具有重要的实用价值。基于细菌群落中普遍存在群感效应的基本原理并结合酶促催化的动力学特征,首先在大肠杆菌群落中建立信号分子高丝氨酸内酯(AHL)介导的细胞–细胞交流机制,将靶基因egfp置入到启动子PluxI的控制之下。在细胞生长过程中,产生的AHL累积到一定浓度启动靶基因表达。通过在细胞生长的不同阶段启动AHL降解酶AiiA的表达控制环境中信号分子AHL的浓度水平,从而控制靶基因egfp的转录效率,最终实现对靶蛋白EGFP表达水平的精确控制。通过检测细胞的生长状态、靶基因在mRNA水平、蛋白质水平的表达情况证明人工设计的分子开关可以便捷高效地控制靶基因表达水平,具有时空调节的严谨性。该分子开关有望广泛应用于代谢工程和合成生物学等研究领域中。 相似文献
77.
构建了新型联合基因载体pcDNA3.1(-)VEGF-siRNA/yCDglyTK,研究其在人胃癌细胞系SGC7901细胞中的表达和杀伤作用.构建靶向血管内皮生长因子(VEGF)的干扰质粒pGenesil-VEGF-siRNA,采用PCR法从中扩增siRNA表达框(含U6启动子),亚克隆至双自杀基因载体pcDNA3.1(-)CV-yCDglyTK,构建联合基因质粒pcDNA3.1(-)VEGF-siRNA/yCDglyTK;通过酶切、测序等鉴定重组质粒;以磷酸钙纳米颗粒为载体,将干扰质粒、双自杀基因质粒及联合基因质粒转染SGC7901细胞,RT-PCR、Western-blot验证目的基因表达;MTT法检测转染细胞对5-氟胞嘧啶(5-FC)的敏感性.结果表明:酶切及测序证实联合基因载体pcDNA3.1(-)VEGF-siRNA/yCDglyTK构建成功;SGC7901细胞转染联合基因质粒后,RT-PCR、Western-blot证实融合自杀基因表达,而VEGF基因表达下调;在前体药物5-FC作用下,转染联合基因组细胞存活率最低,与其他组比较有统计学差异.成功构建联合基因载体pcDNA3.1(-)VEGF-si... 相似文献
78.
79.
Prem Pritam Aditya P. Sarnaik Pramod P. Wangikar 《Biotechnology and bioengineering》2023,120(8):2363-2370
With multiple applications in food, pharmaceutical, and chemical industries as antioxidant or nonmetabolizable sweetener; the bioproduction of d -mannitol is gaining global attention, especially with photosynthetic organisms as hosts. Considering the sustainability prospects, the current work encompasses metabolic engineering of a widely used cyanobacterial strain, Synechococcus elongatus PCC 7942, and two newly isolated fast-growing cyanobacterial strains; S. elongatus PCC 11801 and S. elongatus PCC 11802, for mannitol production. We engineered these strains with a two-step pathway by cloning genes for mannitol-1-phosphate dehydrogenase (mtlD) and mannitol-1-phosphatase (mlp), where the mtlD expression was under the control of different promoters from PCC 7942, namely, Prbc225, PcpcB300, PcpcBm1, PrbcLm17, and PrbcLm15. The strains were tested under the “switch conditions,” where the growth conditions were switched after the first 3 days, thereby resulting in differential promoter activity. Among the engineered strains of PCC 11801 and PCC 11802, the strains possessing Prbc225-mtlD module produced relatively high mannitol titers of 401 ± 18 mg/L and 537 ± 18 mg/L, respectively. The highest mannitol titer of 701 ± 15 mg/L (productivity 60 mg/L.d, yield 895 µM/OD730) was exhibited by the engineered strain of PCC 7942 expressing PcpcB300-mtlD module. It is by far the highest obtained mannitol yield from the engineered cyanobacteria. 相似文献
80.
Abstract. Strophingia ericae (Curtis) (Homoptera: Psylloidea) takes one or two years to complete its life cycle. In both cases eggs hatch over a prolonged period from midsummer, possibly extending into the following spring at high altitude, and instars overlap in time. Instar III is the predominant overwintering stage in the lowland, annual cycle, whereas in the upland, biennial cycle most first-year nymphs overwinter in instars I and II and most second-year nymphs in instar V. When moved to the laboratory, instars IV and V from a predominantly annual population showed accelerated development in response to elevated temperatures and to long days in both mid-winter and early spring. In the biennial life cycle, short autumn days retard instar V development but the response to photoperiod disappears by the end of winter. Exposure to LD 18 : 6 h retarded development of early instars in the annual population, resulting in an accumulation in instar III. The proportion of overwintering instars I and II rises with increase in altitude and moult to instar III becomes progressively delayed. Nymphs that reach instar III under long daylengths in the year following hatching are channelled towards the biennial cycle. Exposure of a predominantly biennial population to 15 °C and LD 18 : 6 h after midsummer, thus avoiding autumn conditions, promoted the rapid development of overwintered nymphs, switching the cycle from biennial to annual. 相似文献