Characterization of bacterial-type phosphoenolpyruvate carboxylase expressed in male gametophyte of higher plants

<p>Abstract</p> <p>Background</p> <p>Phospho<it>enol</it>pyruvate carboxylase (PEPC) is a critical enzyme catalyzing the β-carboxylation of phospho<it>enol</it>pyruvate (PEP) to oxaloacetate, a tricarboxylic acid (TCA) cycle intermediate. PEPC typically exists as a Class-1 PEPC homotetramer composed of plant-type PEPC (PTPC) polypeptides, and two of the subunits were reported to be monoubiquitinated in germinating castor oil seeds. By the large-scale purification of ubiquitin (Ub)-related proteins from lily anther, two types of PEPCs, bacterial-type PEPC (BTPC) and plant-type PEPC (PTPC), were identified in our study as candidate Ub-related proteins. Until now, there has been no information about the properties of the PEPCs expressed in male reproductive tissues of higher plants.</p> <p>Results</p> <p>Expression analyses showed that lily BTPC (LlBTPC) and <it>Arabidopsis </it>BTPC (AtBTPC) were significantly expressed in pollen. The fusion protein AtBTPC-Venus localized in the cytoplasm of the vegetative cell (VC). Both LlBTPC and AtBTPC expression initiated after the last mitosis before pollen germination. Lily PTPC (LlPTPC) and monoubiquitinated LlPTPC (Ub-LlPTPC) remained at constant levels during pollen development. In late bicellular pollen of lily, LlBTPC forms a hetero-octameric Class-2 PEPC complex with LlPTPC to express PEPC activity.</p> <p>Conclusion</p> <p>Our results suggest that an LlBTPC:Ub-LlPTPC:LlPTPC complex is formed in the VC cytoplasm during late pollen development. Both LlBTPC and AtBTPC expression patterns are similar to the patterns of the appearance of storage organelles during pollen development in lily and <it>Arabidopsis</it>, respectively. Therefore, BTPC is thought to accelerate the metabolic flow for the synthesis of storage substances during pollen maturation. Our study provides the first characterization of BTPC in pollen, the male gametophyte of higher plants.</p

Steering bends and changing lanes: the impact of optic flow and road edges on two point steering control

Successful driving involves steering corrections that respond to immediate positional errors whilst also anticipating upcoming changes to the road layout ahead. In popular steering models these tasks are often treated as separate functions using two points: the near region for correcting current errors, and the far region for anticipating future steering requirements. Whilst two-point control models can capture many aspects of driver behaviour, the nature of perceptual inputs to these two ‘points’ remains unclear. Inspired by experiments that solely focused on road-edge information (Land & Horwood, 1995), two-point models have tended to ignore the role of optic flow during steering control. There is recent evidence demonstrating that optic flow should be considered within two-point control steering models (Mole et al., 2016). To examine the impact of optic flow and road edges on two-point steering control we used a driving simulator to selectively and systematically manipulate these components. We removed flow and/or road-edge information from near or far regions of the scene, and examined how behaviours changed when steering along roads where the utility of far-road information varied. Whilst steering behaviours were strongly influenced by the road-edges, there were also clear contributions of optic flow to steering responses. The patterns of steering were not consistent with optic flow simply feeding into two-point control, rather the global optic flow field appeared to support effective steering responses across the time-course of each trajectory

Isolation and identification of ubiquitin-related proteins from Arabidopsis seedlings

The majority of proteins in eukaryotic cells are modified according to highly regulated mechanisms to fulfill specific functions and to achieve localization, stability, and transport. Protein ubiquitination is one of the major post-translational modifications occurring in eukaryotic cells. To obtain the proteomic dataset related to the ubiquitin (Ub)-dependent regulatory system in Arabidopsis, affinity purification with an anti-Ub antibody under native condition was performed. Using MS/MS analysis, 196 distinct proteins represented by 251 distinct genes were identified. The identified proteins were involved in metabolism (23.0%), stress response (21.4%), translation (16.8%), transport (6.7%), cell morphology (3.6%), and signal transduction (1.5%), in addition to proteolysis (16.8%) to which proteasome subunits (14.3%) is included. On the basis of potential ubiquitination-targeting signal motifs, in-gel mobilities, and previous reports, 78 of the identified proteins were classified as ubiquitinated proteins and the rest were speculated to be associated proteins of ubiquitinated proteins. The degradation of three proteins predicted to be ubiquitinated proteins was inhibited by a proteasome inhibitor, suggesting that the proteins were regulated by Ub/proteasome-dependent proteolysis

Phosphoinositide 3-Kinaseγ Controls the Intracellular Localization of CpG to Limit DNA-PKcs-Dependent IL-10 Production in Macrophages

Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG) stimulate innate immune responses. Phosphoinositide 3-kinase (PI3K) has been implicated in CpG-induced immune activation; however, its precise role has not yet been clarified. CpG-induced production of IL-10 was dramatically increased in macrophages deficient in PI3Kγ (p110γ−/−). By contrast, LPS-induced production of IL-10 was unchanged in the cells. CpG-induced, but not LPS-induced, IL-10 production was almost completely abolished in SCID mice having mutations in DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Furthermore, wortmannin, an inhibitor of DNA-PKcs, completely inhibited CpG-induced IL-10 production, both in wild type and p110γ−/− cells. Microscopic analyses revealed that CpG preferentially localized with DNA-PKcs in p110γ−/− cells than in wild type cells. In addition, CpG was preferentially co-localized with the acidic lysosomal marker, LysoTracker, in p110γ−/− cells, and with an early endosome marker, EEA1, in wild type cells. Over-expression of p110γ in Cos7 cells resulted in decreased acidification of CpG containing endosome. A similar effect was reproduced using kinase-dead mutants, but not with a ras-binding site mutant, of p110γ. Thus, it is likely that p110γ, in a manner independent of its kinase activity, inhibits the acidification of CpG-containing endosomes. It is considered that increased acidification of CpG-containing endosomes in p110γ−/− cells enforces endosomal escape of CpG, which results in increased association of CpG with DNA-PKcs to up-regulate IL-10 production in macrophages

An infant incubator exchange criteria based on bacteria acquisition of postnatal two weeks. <Workshop 1>

保育器交換は頻繁に行われるが, その基準や方法についてのエビデンスは確立していない. 我々は, 生後2週間の7名の新生児と児の使用した7台の保育器の細菌培養結果から, 保育器交換の時期について検討した. 方法は, データは当院が実施している入院時からの児の監視培養結果と, 同時期に採取した保育器の細菌培養結果を用い, 事例ごとの検出細菌の変化を検討した. 結果: 出生直後の患児の細菌培養検査は全例で陰性(No Growth)だった. 生後1週間以内の患児と保育器の培養結果はほとんど陰性だった. 生後1週間での保育器交換の必要性は低いことが示された. また患児からと保育器からの検出菌の種類は類似していた. 考察: 無菌状態から細菌を獲得する過程は次のように考察した. 患児が細菌を獲得し, それが保育器に付着する, または保育器に付着した菌が患児に移行している. 保育器交換の主な目的は2つある. ひとつは感染予防, もうひとつは清掃である. 細菌学的には保育器は患児の生活環境の一部であり, 保育器交換を行ってもまた同様の細菌が検出される可能性が高いと考えた. 通常の清拭のみで拭き取りきれない器内の汚染がある場合, 保育器交換が必要である. 患児の生活環境としての清潔の維持, 患児への侵襲, マンパワー, コスト面などを考慮し本当に必要な保育器交換の時期をどこで判断するかが今後の検討課題である.Objects: Infant incubator exchange (IIE) is performed frequently, but the criteria and evidence about the methods have not been established. We review a timing of infant incubator exchange from a viewpoint of early postnatal bacteria acquisition. Methods: We reviewed bacteriological examination results of seven newborn patients and incubators in which they have accomodated. A change of acquired bacteria in each case was examined based on bacterial culture results of a newborn since admission and on simultaneous culture results of an infant incubator. Results: The bacteriological examination of a newborn patient just after birth was negative (no growth) in all cases. Newborn patient of postnatal less than one week and most of an infant incubator were negative in bacterial culture. It was shown that need of IIE in postnatal one week was low. In addition, the kind of cultured bacteria from an infant incubator was similar to it from a patient. Discussions: The process to acquire bacterial flora from aseptic condition was supposed as follows: patient acquires bacteria, and it sticks to an infant incubator, or the bacteria which stuck to an infant incubator move to a patient. There are two main purposes of IIE, one is to prevent infectious diseases, the other is cleaning. An infant incubator was bacteriologically a part of living environment of a patient, and we thought it was very likely that a similar kind of bacteria can be detected even if an IIE was carried out frequently. When you find contamination in the incubator which cannot being wiped off only in a usual method, IIE is required. Keeping sanitary as living environment of a patient, aggression to a patient, manpower, a cost side must be considered, but it remains controversial how and when to decide a timing of really necessary IIE

Comparative Analysis of Depth Detection Algorithms using Stereo Vision

In recent years, the use of unmanned aerial vehicles in various domains has increased exponentially. Drones are being extensively used in the fields of Agriculture, Transportation, military, etc. Different sensors are being integrated into the drones depending upon the application. Lately, LIDAR sensors are being integrated on the drone for acquiring depth-related information. Though these sensors have advantages, they are very costly and do not perform very well under high sun angles. Stereo cameras can be mounted on drones to get depth perception of the obstacles as they are cheaper and efficient. In this paper, we have developed and compared two different algorithms ( Conventional and deep learning-based) for realtime depth detection of the obstacle using stereo vision with the intention to mount the stereo camera on drone in future. The comparison is based on accuracy, range of operation and load incurred by the algorithm on the system. The coefficient of determination (R 2) and correlation coefficient has also been calculated which shows that Algorithm1 exhibits correlation and R 2 value of 0:9985 and 0:9971 respectively. This is considerably higher than Algorithm2 whose values are around 0:8779 and 0:7707 respectively

Impact of Transferrin Saturation and Anemia on Radial Artery Calcification in Patients with End-Stage Kidney Disease

Background: Arterial calcification is an important factor in determining the prognosis of patients with chronic kidney disease (CKD). Few studies on aortic calcification have involved radial artery calcification (RAC). This study aimed to analyze risk factors for RAC in patients with end-stage kidney disease (ESKD) and investigate the relationship between subsequent cardiovascular events (CVE) and vascular access trouble (VAT). Methods: This cohort study included 64 consecutive patients with ESKD who initiated hemodialysis and underwent a procedure for the creation of a primary radiocephalic arteriovenous fistula (RCAVF). Small arterial specimens were obtained from patients during RCAVF surgery. Tissue samples were stained with von Kossa, and arterial microcalcification was evaluated. We analyzed the association between preexisting arterial microcalcifications, clinical characteristics, CVE, and VAT. Results: In the univariate analysis, RAC patients demonstrated high systolic blood pressure (sBP), low hemoglobin (Hb), and low transferrin saturation (TSAT) (&lt;0.05, &lt;0.05, and &lt;0.05, respectively). In the multivariate analysis, Hb (HR&ndash;0.516 (0.278&ndash;0.959), p &lt; 0.05), TSAT (HR&ndash;0.0012 (0.00000248&ndash;0.597), p &lt; 0.05), and sBP (HR&ndash;1.037 (1.001&ndash;1.073), p &lt; 0.05) were independent risk factors for RAC. The cumulative incidence rate of CVE/VAT was not associated with RAC for one year. Conclusion: RAC was associated with sBP, TSAT, and anemia; however, no association with CVE/VAT was observed