Strain Construction and Screening Methods for a Yeast Histone H3/H4 Mutant Library

Author(s):  
Junbiao Dai ◽  
Jef D. Boeke
2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Ran Tu ◽  
Yue Zhang ◽  
Erbing Hua ◽  
Likuan Bai ◽  
Huamei Huang ◽  
...  

AbstractStreptomyces are one of the most important industrial microorganisms for the production of proteins and small-molecule drugs. Previously reported flow cytometry-based screening methods can only screen spores or protoplasts released from mycelium, which do not represent the filamentous stationary phase Streptomyces used in industrial cultivation. Here we show a droplet-based microfluidic platform to facilitate more relevant, reliable and rapid screening of Streptomyces mycelium, and achieved an enrichment ratio of up to 334.2. Using this platform, we rapidly characterized a series of native and heterologous constitutive promoters in Streptomyces lividans 66 in droplets, and efficiently screened out a set of engineered promoter variants with desired strengths from two synthetic promoter libraries. We also successfully screened out several hyperproducers of cellulases from a random S. lividans 66 mutant library, which had 69.2–111.4% greater cellulase production than the wild type. Our method provides a fast, simple, and powerful solution for the industrial engineering and screening of Streptomyces in more industry-relevant conditions.


1978 ◽  
Vol 17 (01) ◽  
pp. 36-40 ◽  
Author(s):  
J.-P. Durbec ◽  
Jaqueline Cornée ◽  
P. Berthezene

The practice of systematic examinations in hospitals and the increasing development of automatic data processing permits the storing of a great deal of information about a large number of patients belonging to different diagnosis groups.To predict or to characterize these diagnosis groups some descriptors are particularly useful, others carry no information. Data screening based on the properties of mutual information and on the log cross products ratios in contingency tables is developed. The most useful descriptors are selected. For each one the characterized groups are specified.This approach has been performed on a set of binary (presence—absence) radiological variables. Four diagnoses groups are concerned: cancer of pancreas, chronic calcifying pancreatitis, non-calcifying pancreatitis and probable pancreatitis. Only twenty of the three hundred and forty initial radiological variables are selected. The presence of each corresponding sign is associated with one or more diagnosis groups.


1964 ◽  
Vol 11 (02) ◽  
pp. 506-512 ◽  
Author(s):  
V. A Lovric ◽  
J Margolis

SummaryAn adaptation of “kaolin clotting time” and prothrombin time for use on haemolysed capillary blood provided simple and sensitive screening tests suitable for use in infants and children. A survey of three year’s experience shows that these are reliable routine laboratory tests for detection of latent coagulation disorders.


1997 ◽  
Vol 78 (05) ◽  
pp. 1360-1365 ◽  
Author(s):  
G Freyburger ◽  
S Javorschi ◽  
S Labrouche ◽  
P Bernard

SummaryThe aim of the present study was to evaluate the relative performance of five screening methods for APC resistance caused by the factor V:Q506 mutation: the original method Coatest® APC™ Resistance Chromogenix, a modified method using the same reagents but a predilution 1+4 of the plasma in a factor V deficient plasma from Stago (Stago deficient V) or from Chromogenix (V-DEF Plasma), the Coatest® APC™ Resistance V (Chromogenix), and Accelerimat™ from bioMérieux. Normalization was done against a pool of normal plasmas for the methods from Chromogenix. The study included 350 subjects, 219 were genotyped (174 FV:R506R, 42 FV:Q506R, 3 FV:Q506Q) and most of them were assessed by more than one method. Uncertainty in predicting the FV genotype was evaluated by statistical analysis, which provided a way to quantitate the performance of the different diagnostic approaches. Performance of each test was evaluated by its sensitivity, specificity, R.O.C. curves, positive and negative likelihood ratios (LR), and the overall performance was determined by two parameters derived from the LR curves : the maximum LR value obtained at the crossover of the two curves, and the distance between the two curves for LR = 10. Coatest® APC™ Resistance V and Accelerimat™ were proven to be the methods most able to discriminate for factor V:Q506, while normalization was not shown to improve the screening performance. The original method from Chromogenix was confirmed to undergo many influences (factor XII, PAI-1, thrombin- antithrombin complexes, antithrombin III, hematocrit). Although a very good improvement was provided by the newest methods, they were shown to be influenced by protein S and/or factor V levels in the sample plasma.


2009 ◽  
Vol 13 (4) ◽  
pp. 119-126
Author(s):  
Yoshimitsu Bamba ◽  
Tetsunari Nishikawa ◽  
Akio Tanaka

2015 ◽  
Vol 24 (2) ◽  
pp. 197-201 ◽  
Author(s):  
Ramesh P. Arasaradnam ◽  
Michael McFarlane ◽  
Emma Daulton ◽  
Erik Westenbrink ◽  
Nicola O’Connell ◽  
...  

Background & Aims: Non-Alcoholic Fatty Liver Disease (NAFLD) is the commonest cause of chronic liver disease in the western world. Current diagnostic methods including Fibroscan have limitations, thus there is a need for more robust non-invasive screening methods. The gut microbiome is altered in several gastrointestinal and hepatic disorders resulting in altered, unique gut fermentation patterns, detectable by analysis of volatile organic compounds (VOCs) in urine, breath and faeces. We performed a proof of principle pilot study to determine if progressive fatty liver disease produced an altered urinary VOC pattern; specifically NAFLD and Non-Alcoholic Steatohepatitis (NASH).Methods: 34 patients were recruited: 8 NASH cirrhotics (NASH-C); 7 non-cirrhotic NASH; 4 NAFLD and 15 controls. Urine was collected and stored frozen. For assay, the samples were defrosted and aliquoted into vials, which were heated to 40±0.1°C and the headspace analyzed by FAIMS (Field Asymmetric Ion Mobility Spectroscopy). A previously used data processing pipeline employing a Random Forrest classification algorithm and using a 10 fold cross validation method was applied.Results: Urinary VOC results demonstrated sensitivity of 0.58 (0.33 - 0.88), but specificity of 0.93 (0.68 - 1.00) and an Area Under Curve (AUC) 0.73 (0.55 -0.90) to distinguish between liver disease and controls. However, NASH/NASH-C was separated from the NAFLD/controls with a sensitivity of 0.73 (0.45 - 0.92), specificity of 0.79 (0.54 - 0.94) and AUC of 0.79 (0.64 - 0.95), respectively.Conclusions: This pilot study suggests that urinary VOCs detection may offer the potential for early non-invasive characterisation of liver disease using 'smell prints' to distinguish between NASH and NAFLD.


2018 ◽  
Vol 11 (2) ◽  
pp. 95-104
Author(s):  
Ivan D. Ivanov ◽  
Stefan A. Buzalov ◽  
Nadezhda H. Hinkova

Summary Preterm birth (PTB) is a worldwide problem with great social significance because it is a leading cause of perinatal complications and perinatal mortality. PTB is responsible for more than a half of neonatal deaths. The rate of preterm delivery varies between 5-18% worldwide and has not decreased in recent years, regardless of the development of medical science. One of the leading causes for that is the failure to identify the high-risk group in prenatal care. PTB is a heterogeneous syndrome in which many different factors interfere at different levels of the pathogenesis of the initiation of delivery, finally resulting in delivery before 37 weeks of gestation (wg). The various specificities of risk factors and the unclear mechanism of initiation of labour make it difficult to elaborate standard, unified and effective screening to diagnose pregnant women at high-risk for PTB correctly. Furthermore, they make primary and secondary prophylaxis less effective and render diagnostic and therapeutic measures ineffective and inappropriate. Reliable and accessible screening methods are necessary for antenatal care, and risk factors for PTB should be studied and clarified in search of useful tools to solve issues of risk pregnancies to decrease PTB rates and associated complications.


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