scholarly journals Integration of a FT expression cassette into CRISPR/Cas9 construct enables fast generation and easy identification of transgene-free mutants in Arabidopsis

2019 ◽  
Author(s):  
Yuxin Cheng ◽  
Na Zhang ◽  
Saddam Hussain ◽  
Sajjad Ahmed ◽  
Wenting Yang ◽  
...  

AbstractThe CRISPR/Cas9 genome editing technique has been widely used to generate transgene-free mutants in different plant species. Several different methods including fluorescence marker-assisted visual screen of transgene-free mutants and programmed self-elimination of CRISPR/Cas9 construct have been use to increase the efficiency of genome edited transgene-free mutants isolation, but the overall time length required to obtain transgene-free mutants has remained unchanged in these methods. We report here a method for fast generation and easy identification of transgene-free mutants in Arabidopsis. By generating and using a single FT expression cassette-containing CRISPR/Cas9 construct, we targeted two sites of the AITR1 gene. We obtained many early bolting plants in T1 generation, and found that about two thirds of these plants have detectable mutations. We then analyzed T2 generations of two representative lines of genome edited early bolting T1 plants, and identified plants without early bolting phenotype, i.e., transgene-free plants, for both lines. Further more, homologues aitr1 mutants were successful obtained for both lines from these transgene-free plants. Taken together, these results suggest that the method described here enables fast generation, and at the mean time, easy identification of transgene-free mutants in plants.


1985 ◽  
Vol 63 (7) ◽  
pp. 1292-1295 ◽  
Author(s):  
Xavier Nesme ◽  
Philippe Normand ◽  
Francine M. Tremblay ◽  
Maurice Lalonde

The question of compatibility between actinorhizal host plants and Frankia sp. was addressed using nodulation speed on Alnus spp. seedlings and Myrica gale seedlings. It was found that the speed of nodulation, defined as the mean time taken for the formation of the first prenodule, was a stable phenotype of both the Frankia strains and the host plants and that a distinction between slow-, medium-, and fast-nodulating Frankia strains could be made. The speed of nodulation of a given Frankia strain did not appear to be positively correlated to the original host plant from which isolation was first performed. It was, however, positively correlated with the Frankia strain and with the host plant species used for inoculation. Some optimal host plant – endophyte combinations were thus defined. Pure spore inocula of Frankia and in vitro propagated Alnus glutinosa plantlets were used to confirm that both the host plant and the microbial partners genetically influenced the nodulation process.



2019 ◽  
Vol 21 (1) ◽  
pp. 49-57 ◽  
Author(s):  
Marek Bąkowski ◽  
Agnieszka Filipiak ◽  
Zdenek Fric

The foraging behaviour of the endangered butterfly Lycaena dispar Haw. was examined in a wet meadow in Poznan (western Poland) in the summer of 2003. Observations showed that the males spent more time resting (11.3% compared to 5.9%) and less time nectaring (24.8% compared 35%) compared to females. The mean time of one visit on a flower was almost three times shorter in males than in females. In total, adults visited flowers of nine nectar plant species, the most frequent ones were Inula britannica, Lychnis flos-cuculi and Cirsium arvense, which were some of the most abundant plant species there. We observed differences of nectar plant use between sexes and generations of the butterfly, but did not confirm preference for the plant colour.



2014 ◽  
Vol 66 (3-4) ◽  
pp. 379-386 ◽  
Author(s):  
Marcello Lisci ◽  
Ettore Pacini

The fruit of <em>Mercurialis annua</em> L. is a two-seeded capsule with a caruncle, a small appendage which arises from the outer integument in the micropylar area. The inner integument has a thick layer of Malpighian-like cells, interrupted at the micropyle by a remnant of nucellus, the walls of which contain suberin. There is no cuticle covering the caruncle and its cells have lipids and protein bodies as reserves. Seed dispersal occurs by a combination of autochory followed by myrmecochory. The explosive ejection of seeds is caused by dehydration-induced torsion of the mechanical layer of the fruit walls. This phenomenon is due to the different orientation of the cells and a folding over of the mechanical layer in the chalazal area. The seeds were found to be thrown distances from 1 to 130 cm (mean 41.1 cm; modal peak 10-20 cm). Ballistic dispersal is influenced by the weight of the seed. The seeds were collected by ants in a mean time of 24.4 minutes the maximum and mean distances of dispersal being 14 and 3.4 m, respectively. Of the seeds removed, 95.2 % were recovered on excavation of the ant nest. The seeds in the nest were intact but without the caruncles. The mean distance of dispersal by diplochory was &lt; 5 m. These features are discussed in relation to possible benefits to the plant species in terms of adaptive advantages of seeds.



1996 ◽  
Vol 75 (05) ◽  
pp. 731-733 ◽  
Author(s):  
V Cazaux ◽  
B Gauthier ◽  
A Elias ◽  
D Lefebvre ◽  
J Tredez ◽  
...  

SummaryDue to large inter-individual variations, the dose of vitamin K antagonist required to target the desired hypocoagulability is hardly predictible for a given patient, and the time needed to reach therapeutic equilibrium may be excessively long. This work reports on a simple method for predicting the daily maintenance dose of fluindione after the third intake. In a first step, 37 patients were delivered 20 mg of fluindione once a day, at 6 p.m. for 3 consecutive days. On the morning of the 4th day an INR was performed. During the following days the dose was adjusted to target an INR between 2 and 3. There was a good correlation (r = 0.83, p<0.001) between the INR performed on the morning of day 4 and the daily maintenance dose determined later by successive approximations. This allowed us to write a decisional algorithm to predict the effective maintenance dose of fluindione from the INR performed on day 4. The usefulness and the safety of this approach was tested in a second prospective study on 46 patients receiving fluindione according to the same initial scheme. The predicted dose was compared to the effective dose soon after having reached the equilibrium, then 30 and 90 days after. To within 5 mg (one quarter of a tablet), the predicted dose was the effective dose in 98%, 86% and 81% of the patients at the 3 times respectively. The mean time needed to reach the therapeutic equilibrium was reduced from 13 days in the first study to 6 days in the second study. No hemorrhagic complication occurred. Thus the strategy formerly developed to predict the daily maintenance dose of warfarin from the prothrombin time ratio or the thrombotest performed 3 days after starting the treatment may also be applied to fluindione and the INR measurement.



2021 ◽  
pp. 107815522110160
Author(s):  
Bernadatte Zimbwa ◽  
Peter J Gilbar ◽  
Mark R Davis ◽  
Srinivas Kondalsamy-Chennakesavan

Purpose To retrospectively determine the rate of death occurring within 14 and 30 days of systemic anticancer therapy (SACT), compare this against a previous audit and benchmark results against other cancer centres. Secondly, to determine if the introduction of immune checkpoint inhibitors (ICI), not available at the time of the initial audit, impacted mortality rates. Method All adult solid tumour and haematology patients receiving SACT at an Australian Regional Cancer Centre (RCC) between January 2016 and July 2020 were included. Results Over a 55-month period, 1709 patients received SACT. Patients dying within 14 and 30 days of SACT were 3.3% and 7.0% respectively and is slightly higher than our previous study which was 1.89% and 5.6%. Mean time to death was 15.5 days. Males accounted for 63.9% of patients and the mean age was 66.8 years. 46.2% of the 119 patients dying in the 30 days post SACT started a new line of treatment during that time. Of 98 patients receiving ICI, 22.5% died within 30 days of commencement. Disease progression was the most common cause of death (79%). The most common place of death was the RCC (38.7%). Conclusion The rate of death observed in our re-audit compares favourably with our previous audit and is still at the lower end of that seen in published studies in Australia and internationally. Cases of patients dying within 30 days of SACT should be regularly reviewed to maintain awareness of this benchmark of quality assurance and provide a feedback process for clinicians.



2021 ◽  
pp. 1-7
Author(s):  
Naomi Vather-Wu ◽  
Matthew D. Krasowski ◽  
Katherine D. Mathews ◽  
Amal Shibli-Rahhal

Background: Expert guidelines recommend annual monitoring of 25-hydroxyvitamin D (25-OHD) and maintaining 25-OHD ≥30 ng/ml in patients with dystrophinopathies. Objective: We hypothesized that 25-OHD remains stable and requires less frequent monitoring in patients taking stable maintenance doses of vitamin D. Methods: We performed a retrospective cohort study, using the electronic health record to identify 26 patients with dystrophinopathies with a baseline 25-OHD ≥30 ng/mL and at least one additional 25-OHD measurement. These patients had received a stable dose of vitamin D for ≥3 months prior to their baseline 25-OHD measurement and throughout follow-up. The main outcome measured was the mean duration time the subjects spent with a 25-OHD ≥30 ng/mL. Results: Only 19% of patients dropped their 25-OHD to <  30 ng/ml, with a mean time to drop of 33 months and a median nadir 25-OHD of 28 ng/mL. Conclusions: These results suggest that measurement of 25-OHD every 2–2.5 years may be sufficient in patients with a baseline 25-OHD ≥30 ng/mL and who are on a stable maintenance dose of vitamin D. Other patients may require more frequent assessments.



Electronics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 876
Author(s):  
Igor Gonçalves ◽  
Laécio Rodrigues ◽  
Francisco Airton Silva ◽  
Tuan Anh Nguyen ◽  
Dugki Min ◽  
...  

Surveillance monitoring systems are highly necessary, aiming to prevent many social problems in smart cities. The internet of things (IoT) nowadays offers a variety of technologies to capture and process massive and heterogeneous data. Due to the fact that (i) advanced analyses of video streams are performed on powerful recording devices; while (ii) surveillance monitoring services require high availability levels in the way that the service must remain connected, for example, to a connection network that offers higher speed than conventional connections; and that (iii) the trust-worthy dependability of a surveillance system depends on various factors, it is not easy to identify which components/devices in a system architecture have the most impact on the dependability for a specific surveillance system in smart cities. In this paper, we developed stochastic Petri net models for a surveillance monitoring system with regard to varying several parameters to obtain the highest dependability. Two main metrics of interest in the dependability of a surveillance system including reliability and availability were analyzed in a comprehensive manner. The analysis results show that the variation in the number of long-term evolution (LTE)-based stations contributes to a number of nines (#9s) increase in availability. The obtained results show that the variation of the mean time to failure (MTTF) of surveillance cameras exposes a high impact on the reliability of the system. The findings of this work have the potential of assisting system architects in planning more optimized systems in this field based on the proposed models.



Trauma ◽  
2021 ◽  
pp. 146040862094972
Author(s):  
Ahmed Fadulelmola ◽  
Rob Gregory ◽  
Gavin Gordon ◽  
Fiona Smith ◽  
Andrew Jennings

Introduction: A novel virus, SARS-CoV-2, has caused a fatal global pandemic which particularly affects the elderly and those with comorbidities. Hip fractures affect elderly populations, necessitate hospital admissions and place this group at particular risk from COVID-19 infection. This study investigates the effect of COVID-19 infection on 30-day hip fracture mortality. Method: Data related to 75 adult hip fractures admitted to two units during March and April 2020 were reviewed. The mean age was 83.5 years (range 65–98 years), and most (53, 70.7%) were women. The primary outcome measure was 30-day mortality associated with COVID-19 infection. Results: The COVID-19 infection rate was 26.7% (20 patients), with a significant difference in the 30-day mortality rate in the COVID-19-positive group (10/20, 50%) compared to the COVID-19-negative group (4/55, 7.3%), with mean time to death of 19.8 days (95% confidence interval: 17.0–22.5). The mean time from admission to surgery was 43.1 h and 38.3 h, in COVID-19-positive and COVID-19-negative groups, respectively. All COVID-19-positive patients had shown symptoms of fever and cough, and all 10 cases who died were hypoxic. Seven (35%) cases had radiological lung findings consistent of viral pneumonitis which resulted in mortality (70% of mortality). 30% ( n = 6) contracted the COVID-19 infection in the community, and 70% ( n = 14) developed symptoms after hospital admission. Conclusion: Hip fractures associated with COVID-19 infection have a high 30-day mortality. COVID-19 testing and chest X-ray for patients presenting with hip fractures help in early planning of high-risk surgeries and allow counselling of the patients and family using realistic prognosis.



2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Nikolaos Halidias

Abstract In this note we study the probability and the mean time for absorption for discrete time Markov chains. In particular, we are interested in estimating the mean time for absorption when absorption is not certain and connect it with some other known results. Computing a suitable probability generating function, we are able to estimate the mean time for absorption when absorption is not certain giving some applications concerning the random walk. Furthermore, we investigate the probability for a Markov chain to reach a set A before reach B generalizing this result for a sequence of sets A 1 , A 2 , … , A k {A_{1},A_{2},\dots,A_{k}} .



Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2342
Author(s):  
Corentin Martens ◽  
Olivier Debeir ◽  
Christine Decaestecker ◽  
Thierry Metens ◽  
Laetitia Lebrun ◽  
...  

Recent works have demonstrated the added value of dynamic amino acid positron emission tomography (PET) for glioma grading and genotyping, biopsy targeting, and recurrence diagnosis. However, most of these studies are based on hand-crafted qualitative or semi-quantitative features extracted from the mean time activity curve within predefined volumes. Voxelwise dynamic PET data analysis could instead provide a better insight into intra-tumor heterogeneity of gliomas. In this work, we investigate the ability of principal component analysis (PCA) to extract relevant quantitative features from a large number of motion-corrected [S-methyl-11C]methionine ([11C]MET) PET frames. We first demonstrate the robustness of our methodology to noise by means of numerical simulations. We then build a PCA model from dynamic [11C]MET acquisitions of 20 glioma patients. In a distinct cohort of 13 glioma patients, we compare the parametric maps derived from our PCA model to these provided by the classical one-compartment pharmacokinetic model (1TCM). We show that our PCA model outperforms the 1TCM to distinguish characteristic dynamic uptake behaviors within the tumor while being less computationally expensive and not requiring arterial sampling. Such methodology could be valuable to assess the tumor aggressiveness locally with applications for treatment planning and response evaluation. This work further supports the added value of dynamic over static [11C]MET PET in gliomas.



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