Direct selection for mutators in Escherichia coli

We have constructed strains that allow a direct selection for mutators of Escherichia coli on a single plate medium. The plate selection is based on using two different markers whose reversion is enhanced by a given mutator. Plates containing limiting amounts of each respective nutrient allow the growth of ghost colonies or microcolonies that give rise to full-size colonies only if a reversion event occurs. Because two successive mutational events are required, mutator cells are favored to generate full-size colonies. Reversion of a third marker allows direct visualization of the mutator phenotype by the large number of blue papillae in the full-size colonies. We also describe plate selections involving three successive nutrient markers followed by a fourth papillation step. Different frameshift or base substitution mutations are used to select for mismatch-repair-defective strains (mutHLS and uvrD). We can detect and monitor mutator cells arising spontaneously, at frequencies lower than 10(-5) in the population. Also, we can measure a mutator cascade, in which one type of mutator (mutT) generates a second mutator (mutHLS) that then allows stepwise frameshift mutations. We discuss the relevance of mutators arising on a single medium as a result of cells overcoming successive growth barriers to the development and progression of cancerous tumors, some of which are mutator cell lines.     

A novel way to induce erythroid progenitor self renewal: cooperation of c-Kit with the erythropoietin receptor

Red blood cells are of vital importance for oxygen transport in vertebrates. Thus, their formation during development and homeostasis requires tight control of both progenitor proliferation and terminal red cell differentiation. Self renewal (i.e. long-term proliferation without differentiation) of committed erythroid progenitors has recently been shown to contribute to this regulation. Avian erythroid progenitors expressing the EGF receptor/c-ErbB (SCF/TGFalpha progenitors) can be induced to long-term proliferation by the c-ErbB ligand transforming growth factor alpha and the steroids estradiol and dexamethasone. These progenitors have not yet been described in mammals and their factor requirements are untypical for adult erythroid progenitors. Here we describe a second, distinct type of erythroid progenitor (EpoR progenitors) which can be established from freshly isolated bone marrow and is induced to self renew by ligands relevant for erythropoiesis, i.e. erythropoietin, stem cell factor, the ligand for c-Kit and the glucocorticoid receptor ligand dexamethasone. Limiting dilution cloning indicates that these EpoR progenitors are derived from normal BFU-E/CFU-E. For a detailed study, mEpoR progenitors were generated by retroviral expression of the murine Epo receptor in bone marrow erythroblasts. These progenitors carry out the normal erythroid differentiation program in recombinant differentiation factors only. We show that mEpoR progenitors are more mature than SCF/TGFalpha progenitors and also do no longer respond to transforming growth factor alpha and estradiol. In contrast they are now highly sensitive to low levels of thyroid hormone, facilitating their terminal maturation into erythrocytes.     

Production of Major Extracellular Enzymes during Lignocellulose Degradation by Two Streptomycetes in Agitated Submerged Culture

Streptomyces viridosporus T7A and S. badius 252 were grown in 1 to 2% (wt/vol) slurry cultures with mineral salts solution containing 0.6% yeast extract and 100/200 mesh ground and extracted corn lignocellulose at 37°C. Enzyme activities rapidly increased in the first 3 to 4 days and then declined and remained at a relatively constant level. Concentrations of endoglucanase and xylanase produced by S. badius were lower than those produced by S. viridosporus. However, the lignin-peroxidase peak concentration was threefold higher than with S. viridosporus and was obtained at 9 to 10 days of incubation. By polyacrylamide gel analysis, it was determined that peroxidases from both species consisted of four enzymes, with only one, the lignin peroxidase, having high activity. A culture pH of 8.5 was preferable for lignocellulose degradation by S. badius.     

Effect of aluminium on the mineral nutrition of rice

A study of the effect of increasing aluminium concentration in a dilute nutrient solution on various aspects of the mineral nutrient uptake by the rice plant has shown that aluminium exerts a stimulation on dry matter production and nutrient uptake until a concentration threshold was reached. The value of this threshold was influenced by nutrient solution composition and cultivar. Its location could be calculated by adjusting to the experimental points a rate law from enzyme kinetics on substrate inhibition curve. On the other hand, total uptake of aluminium and its concentration in the tops was a monotonic function of aluminium concentration in the nutrient solution, the effect of which was greatly enhanced by increased phosphate concentration. A sensitive cultivar accumulated more aluminium than a resistant one.The effect of phosphate on the alleviation of aluminium toxicity was slight in the range of concentration studied.Nitrogen uptake either as ammonium or nitrate nitrogen was clearly influenced by aluminium concentration when its instantaneous value was measured by the technique of the continuously flowing culture solution. The ammonium uptake rate of two cultivars different in sensitivity to aluminium was such that the sensitive variety took up less ammonium and acidified less the culture solution flowing through the root sysstem with a residence time of a few hours.Minor elements concentration in the tops of the rice plants did not seem to be greatly influenced by aluminium with the notable exception of manganese, the uptake of which was clearly depressed by increasing aluminium concentration.Attempts were made at using the speciation of the nutrient solutions with or without aluminium complexation by fluoride in order to rank the various ionic forms of aluminium according to their toxicity. It seems that the well-known result of primary toxicity due to the free Al-ion is also true for rice but that some toxicity is associated with the AlSO₄-ion.     

Sounding out the hidden data: A concise review of deep learning in photoacoustic imaging

The rapidly evolving field of photoacoustic tomography utilizes endogenous chromophores to extract both functional and structural information from deep within tissues. It is this power to perform precise quantitative measurements in vivo—with endogenous or exogenous contrast—that makes photoacoustic tomography highly promising for clinical translation in functional brain imaging, early cancer detection, real-time surgical guidance, and the visualization of dynamic drug responses. Considering photoacoustic tomography has benefited from numerous engineering innovations, it is of no surprise that many of photoacoustic tomography’s current cutting-edge developments incorporate advances from the equally novel field of artificial intelligence. More specifically, alongside the growth and prevalence of graphical processing unit capabilities within recent years has emerged an offshoot of artificial intelligence known as deep learning. Rooted in the solid foundation of signal processing, deep learning typically utilizes a method of optimization known as gradient descent to minimize a loss function and update model parameters. There are already a number of innovative efforts in photoacoustic tomography utilizing deep learning techniques for a variety of purposes, including resolution enhancement, reconstruction artifact removal, undersampling correction, and improved quantification. Most of these efforts have proven to be highly promising in addressing long-standing technical obstacles where traditional solutions either completely fail or make only incremental progress. This concise review focuses on the history of applied artificial intelligence in photoacoustic tomography, presents recent advances at this multifaceted intersection of fields, and outlines the most exciting advances that will likely propagate into promising future innovations.     

The influence of host and bacterial genotype on the development of disseminated disease with Mycobacterium tuberculosis

The factors that govern the development of tuberculosis disease are incompletely understood. We hypothesized that some strains of Mycobacterium tuberculosis (M. tuberculosis) are more capable of causing disseminated disease than others and may be associated with polymorphisms in host genes responsible for the innate immune response to infection. We compared the host and bacterial genotype in 187 Vietnamese adults with tuberculous meningitis (TBM) and 237 Vietnamese adults with uncomplicated pulmonary tuberculosis. The host genotype of tuberculosis cases was also compared with the genotype of 392 cord blood controls from the same population. Isolates of M. tuberculosis were genotyped by large sequence polymorphisms. The hosts were defined by polymorphisms in genes encoding Toll-interleukin 1 receptor domain containing adaptor protein (TIRAP) and Toll-like receptor-2 (TLR-2). We found a significant protective association between the Euro-American lineage of M. tuberculosis and pulmonary rather than meningeal tuberculosis (Odds ratio (OR) for causing TBM 0.395, 95% confidence intervals (C.I.) 0.193-0.806, P = 0.009), suggesting these strains are less capable of extra-pulmonary dissemination than others in the study population. We also found that individuals with the C allele of TLR-2 T597C allele were more likely to have tuberculosis caused by the East-Asian/Beijing genotype (OR = 1.57 [95% C.I. 1.15-2.15]) than other individuals. The study provides evidence that M. tuberculosis genotype influences clinical disease phenotype and demonstrates, for the first time, a significant interaction between host and bacterial genotypes and the development of tuberculosis.     

Recognizing brain activities by functional near-infrared spectroscope signal analysis

Background

Noninvasive recording of movements caused by the heartbeat and the blood circulation is known as ballistocardiography. Several studies have shown the capability of a force plate to detect cardiac activity in the human body. The aim of this paper is to present a new method based on differential geometry of curves to handle multivariate time series obtained by ballistocardiographic force plate measurements.

Results

We show that the recoils of the body caused by cardiac motion and blood circulation provide a noninvasive method of displaying the motions of the heart muscle and the propagation of the pulse wave along the aorta and its branches. The results are compared with the data obtained invasively during a cardiac catheterization. We show that the described noninvasive method is able to determine the moment of a particular heart movement or the time when the pulse wave reaches certain morphological structure.

Conclusions

Monitoring of heart movements and pulse wave propagation may be used e.g. to estimate the aortic pulse wave velocity, which is widely accepted as an index of aortic stiffness with the application of predicting risk of heart disease in individuals. More extended analysis of the method is however needed to assess its possible clinical application.     

Design and Application of a New Cryptic-Plasmid-Based Shuttle Vector for Magnetospirillum magneticum

A 3.7-kb cryptic plasmid designated pMGT was found in Magnetospirillum magneticum MGT-1. It was characterized and used for the development of an improved expression system in strain AMB-1 through the construction of a shuttle vector, pUMG. An electroporation method for magnetic bacteria that uses the cryptic plasmid was also developed.