Reciprocal regulation of protein synthesis and carbon metabolism for thylakoid membrane biogenesis

Metabolic control of gene expression coordinates the levels of specific gene products to meet cellular demand for their activities. This control can be exerted by metabolites acting as regulatory signals and/or a class of metabolic enzymes with dual functions as regulators of gene expression. However, little is known about how metabolic signals affect the balance between enzymatic and regulatory roles of these dual functional proteins. We previously described the RNA binding activity of a 63 kDa chloroplast protein from Chlamydomonas reinhardtii, which has been implicated in expression of the psbA mRNA, encoding the D1 protein of photosystem II. Here, we identify this factor as dihydrolipoamide acetyltransferase (DLA2), a subunit of the chloroplast pyruvate dehydrogenase complex (cpPDC), which is known to provide acetyl-CoA for fatty acid synthesis. Analyses of RNAi lines revealed that DLA2 is involved in the synthesis of both D1 and acetyl-CoA. Gel filtration analyses demonstrated an RNP complex containing DLA2 and the chloroplast psbA mRNA specifically in cells metabolizing acetate. An intrinsic RNA binding activity of DLA2 was confirmed by in vitro RNA binding assays. Results of fluorescence microscopy and subcellular fractionation experiments support a role of DLA2 in acetate-dependent localization of the psbA mRNA to a translation zone within the chloroplast. Reciprocally, the activity of the cpPDC was specifically affected by binding of psbA mRNA. Beyond that, in silico analysis and in vitro RNA binding studies using recombinant proteins support the possibility that RNA binding is an ancient feature of dihydrolipoamide acetyltransferases. Our results suggest a regulatory function of DLA2 in response to growth on reduced carbon energy sources. This raises the intriguing possibility that this regulation functions to coordinate the synthesis of lipids and proteins for the biogenesis of photosynthetic membranes

Occurrence and formation of indole-3-acetamide in Arabidopsis thaliana

An HPLC/GC–MS/MS technique (high-pressure liquid chromatography in combination with gas chromatography–tandem mass spectrometry) has been worked out to analyze indole-3-acetamide (IAM) with very high sensitivity, using isotopically labelled IAM as an internal standard. Using this technique, the occurrence of IAM in sterile-grown Arabidopsis thaliana (L.) Heynh. was demonstrated unequivocally. In comparison, plants grown under non-sterile conditions in soil in a greenhouse showed approximately 50% higher average levels of IAM, but the differences were not statistically significant. Thus, microbial contributions to the IAM extracted from the tissue are likely to be minor. Levels of IAM in sterile-grown seedlings were highest in imbibed seeds and then sharply declined during the first 24 h of germination and further during early seedling development to remain below 20–30 pmol g–1 fresh weight throughout the rosette stage. The decline in indole-3-aetic acid (IAA) levels during germination was paralleled by a similar decline in IAM levels. Recombinant nitrilase isoforms 1, 2 and 3, known to synthesize IAA from indole-3-acetonitrile, were shown to produce significant amounts of IAM in vitro as a second end product of the reaction besides IAA. NIT2 was earlier shown to be highly expressed in developing and in mature A. thaliana embryos, and NIT3 is the dominantly active gene in the hypocotyl and the cotyledons of young, germinating seedlings. Collectively, these data suggest that the elevated levels of IAM in seeds and germinating seedlings result from nitrilase action on indole-3-acetonitrile, a metabolite produced in the plants presumably from glucobrassicin turnover

Effect of an interprofessional care concept on the hospitalization of nursing home residents : study protocol for a cluster-randomized controlled trial

Background: The rising number of nursing home (NH) residents and their increasingly complex treatment needs pose a challenge to the German health care system. In Germany, there is no specialized geriatric medical care for NH residents. Nursing staff and general practitioners (GPs) in particular have to compensate for the additional demand, which is compounded by organizational and structural hurdles. As a result, avoidable emergency calls and hospital admissions occur. In the SaarPHIR project (Saarländische PflegeHeimversorgung Integriert Regelhaft), a complex intervention focusing on a medical care concept was developed in a participatory practice-based approach involving NH representatives and GPs. The complex intervention addresses the collaboration between nurses and GPs and aims to help restructure and optimize the existing daily care routine. It is expected to improve the medical care of geriatric patients in NHs and reduce stressful, costly hospital admissions. The intervention was pilot-tested during the first 12 months of the project. In the present study, its effectiveness, cost-effectiveness, and safety will be evaluated. Methods: The study is a cluster-randomized controlled trial, comparing an intervention group with a control group. The intervention includes a concept of interprofessional collaboration, in which GPs group into regional cooperating teams. Teams are encouraged to cooperate more closely with NH staff and to provide on-call schedules, pre-weekend visits, joint team meetings, joint documentation, and improved medication safety. At least 32 NHs in Saarland, Germany (with at least 50 residents each) will be included and monitored for 12 months. The primary endpoint is hospitalization. Secondary endpoints are quality of life, quality of care, and medication safety. The control group receives treatment as usual. Process evaluation and health economic evaluation accompany the study. The data set contains claims data from German statutory health insurance companies as well as primary data. Analysis will be conducted using a generalized linear mixed model. Conclusion: A reduction in hospital admissions of NH residents and relevant changes in secondary endpoints are expected. In turn, these will have a positive impact on the economic assessment

Baseline characteristics of patients in the reduction of events with darbepoetin alfa in heart failure trial (RED-HF)

<p>Aims: This report describes the baseline characteristics of patients in the Reduction of Events with Darbepoetin alfa in Heart Failure trial (RED-HF) which is testing the hypothesis that anaemia correction with darbepoetin alfa will reduce the composite endpoint of death from any cause or hospital admission for worsening heart failure, and improve other outcomes.</p> <p>Methods and results: Key demographic, clinical, and laboratory findings, along with baseline treatment, are reported and compared with those of patients in other recent clinical trials in heart failure. Compared with other recent trials, RED-HF enrolled more elderly [mean age 70 (SD 11.4) years], female (41%), and black (9%) patients. RED-HF patients more often had diabetes (46%) and renal impairment (72% had an estimated glomerular filtration rate <60 mL/min/1.73 m2). Patients in RED-HF had heart failure of longer duration [5.3 (5.4) years], worse NYHA class (35% II, 63% III, and 2% IV), and more signs of congestion. Mean EF was 30% (6.8%). RED-HF patients were well treated at randomization, and pharmacological therapy at baseline was broadly similar to that of other recent trials, taking account of study-specific inclusion/exclusion criteria. Median (interquartile range) haemoglobin at baseline was 112 (106–117) g/L.</p> <p>Conclusion: The anaemic patients enrolled in RED-HF were older, moderately to markedly symptomatic, and had extensive co-morbidity.</p&gt

Chlorella viruses contain genes encoding a complete polyamine biosynthetic pathway

Two genes encoding the putative polyamine biosynthetic enzymes agmatine iminohydrolase (AIH) and N-carbamoylputrescine amidohydrolase (CPA) were cloned from the chloroviruses PBCV-1, NY-2A and MT325. They were expressed in Escherichia coli to form C-terminal (His)6-tagged proteins and the recombinant proteins were purified by Ni2+- binding affinity chromatography. The biochemical properties of the two enzymes are similar to AIH and CPA enzymes from Arabidopsis thaliana and Pseudomonas aeruginosa. Together with the previously known virus genes encoding ornithine/arginine decarboxlyase (ODC/ADC) and homospermidine synthase, the chloroviruses have genes that encode a complete set of functional enzymes that synthesize the rare polyamine homospermidine from arginine via agmatine, N-carbamoylputrescine and putrescine. The PBCV-1 aih and cpa genes are expressed early during virus infection together with the odc/adc gene, suggesting that biosynthesis of putrescine is important in early stages of viral replication. The aih and cpa genes are widespread in the chlorella viruses

Identification and characterization of plant agmatine iminohydrolase, the last missing link in polyamine biosynthesis of plants

The cloning, expression and characterization of plant agmatine iminohydrolase (AIH, also known as agmatine deiminase, EC 3.5.3.12) is described. Recombinant AIH of Arabidopsis thaliana forms dimers and catalyzes the specific conversion of agmatine to N-carbamoylputrescine and ammonia. Biochemical data suggested that cysteine side chains are involved in catalysis. However, site-directed mutagenesis of the two highly conserved cysteine residues of AIH showed that these cysteines are important but not essential for activity, arguing against a thioester substrate-enzyme intermediate during catalysis. This work represents the completion of the cloning of the arginine decarboxylase pathway genes of higher plants. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies

Przegląd metod zapewniających odporność na błędy dla głębokich sieci neuronowych

Asignificant rise in Artificial Intelligence (AI) has impacted many applications around us, so much so that AI has now been increasingly used in safety-critical applications. AI at the edge is the reality, which means performing the data computation closer to the source of the data, as opposed to performing it on the cloud. Safety-critical applications have strict reliability requirements; therefore, it is essential that AI models running on the edge (i.e., hardware) must fulfill the required safety standards. In the vast field of AI, Deep Neural Networks (DNNs) are the focal point of this survey as it has continued to produce extraordinary outcomes in various applications i.e. medical, automotive, aerospace, defense, etc. Traditional reliability techniques for DNNs implementation are not always practical, as they fail to exploit the unique characteristics of the DNNs. Furthermore, it is also essential to understand the targeted edge hardware because the impact of the faults can be different in ASICs and FPGAs. Therefore, in this survey, first, we have examined the impact of the fault in ASICs and FPGAs, and then we seek to provide a glimpse of the recent progress made towards the fault-tolerant DNNs. We have discussed several factors that can impact the reliability of the DNNs. Further, we have extended this discussion to shed light on many state-of-the-art fault mitigation techniques for DNNs.Znaczący rozwój sztucznej inteligencji (SI) wpływa na wiele otaczających nas aplikacji, do tego stopnia, że SI jest obecnie coraz częściej wykorzystywana w aplikacjach o krytycznym znaczeniu dla bezpieczeństwa. Sztuczna inteligencja na brzegu sieci (Edge) jest rzeczywistością, co oznacza wykonywanie obliczeń na danych bliżej źródła danych, w przeciwieństwie do wykonywania ich w chmurze. Aplikacje o krytycznym znaczeniu dla bezpieczeństwa mają wysokie wymagania dotyczące niezawodności; dlatego ważne jest, aby modele SI działające na brzegu sieci (tj. sprzęt) spełniały wymagane standardy bezpieczeństwa. Z rozległej dziedziny sztucznej inteligencji, głębokie sieci neuronowe (DNN) są centralnym punktem tego badania, ponieważ nadal przynoszą znakomite wyniki w różnych zastosowaniach, tj. medycznych, motoryzacyjnych, lotniczych, obronnych itp. Tradycyjne techniki niezawodności implementacji w przypadku DNN nie zawsze są praktyczne, ponieważ nie wykorzystują unikalnych cech DNN. Co więcej, istotne jest również zrozumienie docelowego sprzętu brzegowego, ponieważ wpływ usterek może być różny w układach ASIC i FPGA. Dlatego też w niniejszym przeglądzie najpierw zbadaliśmy wpływ usterek w układach ASIC i FPGA, a następnie staramy się zapewnić wgląd w ostatnie postępy poczynione w kierunku DNN odpornych na błędy. Omówiliśmy kilka czynników, które mogą wpływać na niezawodność sieci DNN. Ponadto rozszerzyliśmy tę dyskusję, aby rzucić światło na wiele najnowocześniejszych technik ograniczania błędów w sieciach DNN