CRISPRi enables fast growth followed by stable aerobic pyruvate formation in Escherichia coli without auxotrophy

CRISPR interference (CRISPRi) was applied to enable the aerobic production of pyruvate in Escherichia coli MG1655 under glucose excess conditions by targeting the promoter regions of aceE or pdhR. Knockdown strains were cultivated in aerobic shaking flasks and the influence of inducer concentration and different sgRNA binding sites on the production of pyruvate was measured. Targeting the promoter regions of aceE or pdhR triggered pyruvate production during the exponential phase and reduced expression of aceE. In lab‐scale bioreactor fermentations, an aceE silenced strain successfully produced pyruvate under fully aerobic conditions during the exponential phase, but loss of productivity occurred during a subsequent nitrogen‐limited phase. Targeting the promoter region of pdhR enabled pyruvate production during the growth phase of cultivations, and a continued low‐level accumulation during the nitrogen‐limited production phase. Combinatorial targeting of the promoter regions of both aceE and pdhR in E. coli MG1655 pdCas9 psgRNA_aceE_234_pdhR_329 resulted in the stable aerobic production of pyruvate with non‐growing cells at YP/S  =  0.36 ± 0.029 gPyruvate/gGlucose in lab‐scale bioreactors throughout an extended nitrogen‐limited production phase

Enantioseparation of Dansylated Amino Acids by Ligand-exchange Capillary Electrophoresis Using L-phenylalaninamide, L-lysine or L-threonine as Chiral Selector

In recent years enantioseparation of both active pharmaceutical ingredients and bio molecules such as amino acids became more and more necessary because in most cases the two stereo forms exhibit different pharmacological effects. This article deals with the chiral separation of dansylated amino acids by ligand-exchange capillary electrophoresis using L-phenylalaninamide, L-lysine and L-threonine as chiral selectors. Experiments with different central metal ions such as Cu(II), Co(II), Cd(II), Ni(II) and Zn(II) were carried out. Optimal conditions were found out by studying the effect of the pH and the selector molarity on the chiral resolution. Best separation was obtained for the Cu(II)/L-lysine complex, showing a chiral resolution up to 17 for Dns-DL-Met. (doi: 10.5562/cca1762

Functional lesional neurosurgery for tremor: back to the future?

For nearly a century, functional neurosurgery has been applied in the treatment of tremor. While deep brain stimulation has been in the focus of academic interest in recent years, the establishment of incisionless technology, such as MRI-guided high-intensity focused ultrasound, has again stirred interest in lesional approaches.In this article, we will discuss the historical development of surgical technique and targets, as well as the technological state-of-the-art of conventional and incisionless interventions for tremor due to Parkinson's disease, essential and dystonic tremor and tremor related to multiple sclerosis (MS) and midbrain lesions. We will also summarise technique-inherent advantages of each technology and compare their lesion characteristics. From this, we identify gaps in the current literature and derive future directions for functional lesional neurosurgery, in particularly potential trial designs, alternative targets and the unsolved problem of bilateral lesional treatment. The results of a systematic review and meta-analysis of the consistency, efficacy and side effect rate of lesional treatments for tremor are presented separately alongside this article

Focused ultrasound ablation as tremor treatment

BACKGROUND: The development of high-intensity magnetic resonance imaging (MRI)-guided focused ultrasound (MRIgFUS) ablation has widened the spectrum of interventional techniques for stereotactic functional neurosurgery of lesions. This has resulted in novel incisionless intervention approaches for the therapy of tremor disorders. The safety and efficacy is documented by recent study data. OBJECTIVES: This article encompasses a description of the technological basis and typical course of MRIgFUS interventions, a comparison to alternative open or incisionless surgical techniques as well as a review of the current evidence base for MRIgFUS ablation in the context of lesional interventions to treat tremor. MATERIAL AND METHODS: Narrative literature review and comparison. METHODS: RESULTS: Depending on the surgical target and tremor etiology published trials of MRIgFUS ablation report a reduction of tremor intensity of up to 80% after 6–12 months follow-up without the disadvantages of open brain surgery. CONCLUSION: The MRIgFUS functional neurosurgery is conducted only at a limited number of treatment sites. First data on lesions of the thalamic ventral intermediary nucleus (V.im.) as well as subthalamic fiber tracts have been published. These results indicate an effective and safe treatment of tremor disorders by MRIgFUS ablation. Incisionless lesional surgery using MRIgFUS is a significant addition to the interventional armamentarium for functional stereotactic neurosurgery and a potentially valuable alternative to established interventional therapy options for tremor disorders

Motility of human renal cells is disturbed by infection with pathogenic hantaviruses

Background: Hemorrhagic fever with renal syndrome (HFRS) caused by pathogenic hantaviruses in Europe and Asia is often characterized by acute kidney injury (AKI) with massive proteinuria. Renal filtration depends on the integrity of epithelial and endothelial monolayers in the tubular and glomerular apparatus. Tubular and glomerular cells represent target cells of hantavirus infection. However, the detailed mechanisms of renal impairment induced by hantaviruses are not well understood. Methods: We analyzed the cellular consequences of hantavirus infection by measuring adhesion and migration capacity of human renal cells infected with Puumala (PUUV) or Hantaan (HTNV) virus. The impact of hantaviral nucleocapsid proteins (N proteins) on motility was examined by transfection of podocytes. Results: Infection of kidney cells with hantavirus species PUUV and HTNV causes a significant reduction of migration capacity. The impaired motility depends on viral replication and transfection of podocytes with N protein of PUUV or HTNV reveals that the expression of N protein alone is sufficient to deteriorate podocyte function. The cellular effects are more pronounced for the more pathogenic HTNV than for PUUV that causes a milder form of HFRS. Conclusions: The direct impairment of migration capacity of renal cells by hantaviral N proteins may contribute substantially to proteinuria observed in the clinical picture of hantavirus infection

Spin sensitive bleaching and monopolar spin orientation in quantum wells

Spin sensitive bleaching of the absorption of far-infrared radiation has been observed in $p$-type GaAs/AlGaAs quantum well structures. The absorption of circularly polarized radiation saturates at lower intensities than that of linearly polarized light due to monopolar spin orientation in the first heavy hole subband. Spin relaxation times of holes in $p$-type material in the range of tens of ps were derived from the intensity dependence of the absorption.Comment: Figures have been updated due to technical printing problems (Postscript mismatch

Spin-polarized Zener tunneling in (Ga,Mn)As

We investigate spin-polarized inter-band tunneling through measurement of (Ga,Mn)As based Zener tunnel diode. By placing the diode under reverse bias, electron spin polarization is transferred from the valence band of p-type (Ga,Mn)As to the conduction band of an adjacent n-GaAs layer. The resulting current is monitored by injection into a quantum well light emitting diode whose electroluminescence polarization is found to track the magnetization of the (Ga,Mn)As layer as a function of both temperature and magnetic field.Comment: 11 pages, 4 figures. Submitted, Physical Review B15 Rapid Communication

An integrative approach to ancient agricultural terraces and forms of dependency: the case of Cutamalla in the prehispanic Andes

This paper presents an integrative and interdisciplinary approach to the study of ancient agricultural terraces and food production systems. Our approach consists of (1) a resource dependency theoretical framework and (2) the application of a variety of archaeological and geoscientific methods, including archaeological and geomorphological surveys, archaeological excavations, drone surveys, mapping based on satellite imagery and high-resolution digital elevation models (DEMs), geographic information system (GIS) applications, soil testing, phytolith analysis, radiocarbon dating, and calculations of food supply capacity and labor requirements. We apply these to the prehispanic site of Cutamalla (3,300 m asl) in the southern Peruvian Andes, which serves as an ideal and pioneering case study. Previous research has focused primarily on the settlement of Cutamalla, particularly through large-scale archaeological excavations, but less attention has been paid to the extensive farming terraces surrounding the settlement and the close relationship between agricultural and settlement activities. By analyzing both the terrace and settlement levels, we take a new perspective and introduce the term agricultural terrace-settlement system for such complexes. Our results show that the residential occupation of Cutamalla and the use of the surrounding farming terraces coincided: the agricultural terrace-settlement system was intensively used for a relatively short period of about 200 years (~250–40 BCE) during the Formative Late Paracas and transitional Initial Nasca periods, long before the famous Inka terrace agricultural systems. There is no evidence of reoccupation of the site and subsequent reuse of the agricultural system. Our data also document the large extent of agricultural terraces around Cutamalla (221 ha) and that maize was likely a major crop grown there. Finally, we place these findings in their broader socio-economic and ecological context. Cutamalla was an important regional center and economic hub during a very dynamic period characterized by significant population growth and increased violence. Not only a more humid climate, but probably also forced collective labor were cornerstones of substantial agricultural production in Cutamalla and the region

Spin dynamics in semiconductors

This article reviews the current status of spin dynamics in semiconductors which has achieved a lot of progress in the past years due to the fast growing field of semiconductor spintronics. The primary focus is the theoretical and experimental developments of spin relaxation and dephasing in both spin precession in time domain and spin diffusion and transport in spacial domain. A fully microscopic many-body investigation on spin dynamics based on the kinetic spin Bloch equation approach is reviewed comprehensively.Comment: a review article with 193 pages and 1103 references. To be published in Physics Reports

RNA-Seq reveals large quantitative differences between the transcriptomes of outbreak and non-outbreak locusts

Outbreaks of locust populations repeatedly devastate economies and ecosystems in large parts of the world. The consequent behavioural shift from solitarious to gregarious and the concomitant changes in the locusts’ biology are of relevant scientific interest. Yet, research on the main locust species has not benefitted from recent advances in genomics. In this first RNA-Seq study on Schistocerca gregaria, we report two transcriptomes, including many novel genes, as well as differential gene expression results. In line with the large biological differences between solitarious and gregarious locusts, almost half of the transcripts are differentially expressed between their central nervous systems. Most of these transcripts are over-expressed in the gregarious locusts, suggesting positive correlations between the levels of activity at the population, individual, tissue and gene expression levels. We group these differentially expressed transcripts by gene function and highlight those that are most likely to be associated with locusts’ phase change either in a species-specific or general manner. Finally, we discuss our findings in the context of population-level and physiological events leading to gregariousness.M. Bakkali wishes to thank the Spanish Ministerio de Ciencia y Tecnología for the for the Ramón y Cajal fellowship and for the BFU2010-16438 grant that supported both this research and the FPI studentship to Rubén Martín Blázquez. We thank Mrs. Pernille Lavgesen for revision of the English language writing of this manuscript. We also thank the editor for the valuable comments on the manuscript