Partial Reduction in BRCA1 Gene Dose Modulates DNA Replication Stress Level and Thereby Contributes to Sensitivity or Resistance

BRCA1 is a well-known breast cancer risk gene, involved in DNA damage repair via homologous recombination (HR) and replication fork protection. Therapy resistance was linked to loss and amplification of the BRCA1 gene causing inferior survival of breast cancer patients. Most studies have focused on the analysis of complete loss or mutations in functional domains of BRCA1. How mutations in non-functional domains contribute to resistance mechanisms remains elusive and was the focus of this study. Therefore, clones of the breast cancer cell line MCF7 with indels in BRCA1 exon 9 and 14 were generated using CRISPR/Cas9. Clones with successful introduced BRCA1 mutations were evaluated regarding their capacity to perform HR, how they handle DNA replication stress (RS), and the consequences on the sensitivity to MMC, PARP1 inhibition, and ionizing radiation. Unexpectedly, BRCA1 mutations resulted in both increased sensitivity and resistance to exogenous DNA damage, despite a reduction of HR capacity in all clones. Resistance was associated with improved DNA double-strand break repair and reduction in replication stress (RS). Lower RS was accompanied by increased activation and interaction of proteins essential for the S phase-specific DNA damage response consisting of HR proteins, FANCD2, and CHK1.     

Circulating miR-200 Family and CTCs in Metastatic Breast Cancer before,during, and after a New Line of Systemic Treatment

The extracellular circulating microRNA (miR)-200 regulates epithelial-mesenchymal transition and, thus, plays an essential role in the metastatic cascade and has shown itself to be a promising prognostic and predictive biomarker in metastatic breast cancer (MBC). Expression levels of the plasma miR-200 family were analyzed in relationship to systemic treatment, circulating tumor cells (CTC) count, progression-free survival (PFS), and overall survival (OS). Expression of miR-200a, miR-200b, miR-200c, miR-141, and miR-429, and CTC status (CTC-positive ≥ 5 CTC/7.5 mL) was assessed in 47 patients at baseline (BL), after the first completed cycle of a new line of systemic therapy (1C), and upon the progression of disease (PD). MiR-200a, miR-200b, and miR-141 expression was reduced at 1C compared to BL. Upon PD, all miR-200s were upregulated compared to 1C. At all timepoints, the levels of miR-200s were elevated in CTC-positive versus CTC-negative patients. Further, heightened miR-200s expression and positive CTC status were associated with poorer OS at BL and 1C. In MBC patients, circulating miR-200 family members decreased after one cycle of a new line of systemic therapy, were elevated during PD, and were indicative of CTC status. Notably, increased levels of miR-200s and elevated CTC count correlated with poorer OS and PFS. As such, both are promising biomarkers for optimizing the clinical management of MBC.     

Mathematical model of the regenerator of a catalytic cracker unit

Conclusions It has been shown that the mathematical model worked out for an industrial regenerator yields satisfactory agreement between the calculated and experimental data on the coke content of the catalyst, the oxygen content in the gas, and the temperatures in the various cross-sections of the regenerator.The best value of the preexponential factor of the regeneration rate constant in an industrial apparatus has been determined.Translated from Khimiya i Tekhnologiya Topliv i Masel, No. 8, pp. 41–43, August, 1967.     

Robotic weed control using automated weed and crop classification

Autonomous robotic weeding systems in precision farming have demonstrated their full potential to alleviate the current dependency on agrochemicals such as herbicides and pesticides, thus reducing environmental pollution and improving sustainability. However, most previous works require fast and constant‐time weed detection systems to achieve real‐time treatment, which forecloses the implementation of more capable but time‐consuming algorithms, for example, learning‐based methods. In this paper, a nonoverlapping multicamera system is applied to provide flexibility for the weed control system in dealing with the indeterminate classification delays. The design, implementation, and testing of our proposed modular weed control unit with mechanical and chemical weeding tools are presented. A framework that performs naive Bayes filtering, 3D direct intra‐ and inter‐camera visual tracking, and predictive control, while integrating state‐of‐the‐art crop/weed detection algorithms, is developed to guide the tools to achieve high‐precision weed removal. The experimental results show that our proposed fully operational weed control system is capable of performing selective mechanical as well as chemical in‐row weeding with indeterminate detection delays in different terrain conditions and crop growth stages.     

Concept and demonstration of an intermediate band tandem device for solar energy conversion

We have realized a tandem solar cell design that combines a pin‐junction with a photovoltaic intersubband absorber. This concept allows harvesting light in the visible range and the near‐ and mid‐infrared at the same time, and theoretically, energy conversion efficiencies beyond the Shockley–Queisser‐limit could be achieved. A test structure was grown, and the operation of this concept could be confirmed, in principal with an optical two‐beam experiment. The basic characteristics of the device can be explained with an equivalent circuit design that consists of three individual cells, and we find an obvious analogy to the concept of the intermediate band solar cell with noteworthy advantages at some points. Our results show, that for a working device it is crucial to adjust the properties of the photovoltaic intersubband absorber for optimal charge separating performance at the working point of the solar cell. Copyright © 2015 John Wiley & Sons, Ltd.     

Electronic structure and chemical bond in stoichiometric hydrides of the group Vb metals

The electronic structure of stoichiometric hydrides of the group Vb metals (V, Nb, Ta) has been investigated by means of the self-consistent full potential scalar-relativistic augmented plane wave method (FP-APW).

The theoretical study of the heat of solution of the ordered monohydrides VH, NbH, and TaH leads to good agreement with experimental values and allows the prediction of the preferred coordination of the hydrogen atoms in these phases, i.e. the octahedral coordination for the VH-system and the tetrahedral coordination for NbH and TaH. Analysis of the charge distribution according to the irreducible representations of the point groups yields a simple orbital picture of the bonding mechanism in the stoichiometric hydride phase. The use of an angular-dependent rather than a spherical-symmetric potential results in a distinct increase in the calculated covalent contribution to the chemical bond.     

Evidence of two oxidative reaction steps initiating anaerobic degradation of resorcinol (1,3-dihydroxybenzene) by the denitrifying bacterium Azoarcus anaerobius

The denitrifying bacterium Azoarcus anaerobius LuFRes1 grows anaerobically with resorcinol (1,3-dihydroxybenzene) as the sole source of carbon and energy. The anaerobic degradation of this compound was investigated in cell extracts. Resorcinol reductase, the key enzyme for resorcinol catabolism in fermenting bacteria, was not present in this organism. Instead, resorcinol was hydroxylated to hydroxyhydroquinone (HHQ; 1,2,4-trihydroxybenzene) with nitrate or K3Fe(CN)6 as the electron acceptor. HHQ was further oxidized with nitrate to 2-hydroxy-1,4-benzoquinone as identified by high-pressure liquid chromatography, UV/visible light spectroscopy, and mass spectroscopy. Average specific activities were 60 mU mg of protein-1 for resorcinol hydroxylation and 150 mU mg of protein-1 for HHQ dehydrogenation. Both activities were found nearly exclusively in the membrane fraction and were only barely detectable in extracts of cells grown with benzoate, indicating that both reactions were specific for resorcinol degradation. These findings suggest a new strategy of anaerobic degradation of aromatic compounds involving oxidative steps for destabilization of the aromatic ring, different from the reductive dearomatization mechanisms described so far.     

Negative regulation of Raf-1 by phosphorylation of serine 621

The elevation of cyclic AMP (cAMP) levels in the cell downregulates the activity of the Raf-1 kinase. It has been suggested that this effect is due to the activation of cAMP-dependent protein kinase (PKA), which can directly phosphorylate Raf-1 in vitro. In this study, we confirmed this hypothesis by coexpressing Raf-1 with the constitutively active catalytic subunit of PKA, which could fully reproduce the inhibition previously achieved by cAMP. PKA-phosphorylated Raf-1 exhibits a reduced affinity for GTP-loaded Ras as well as impaired catalytic activity. As the binding to GTP-loaded Ras induces Raf-1 activation in the cell, we examined which mechanism is required for PKA-mediated Raf-1 inhibition in vivo. A Raf-1 point mutant (RafR89L), which is unable to bind Ras, as well as the isolated Raf-1 kinase domain were still fully susceptible to inhibition by PKA, demonstrating that the phosphorylation of the Raf-1 kinase suffices for inhibition. By the use of mass spectroscopy and point mutants, PKA phosphorylation site was mapped to a single site in the Raf-1 kinase domain, serine 621. Replacement of serine 621 by alanine or cysteine or destruction of the PKA consensus motif by changing arginine 618 resulted in the loss of catalytic activity. Notably, a mutation of serine 619 to alanine did not significantly affect kinase activity or regulation by activators or PKA. Changing serine 621 to aspartic acid yielded a Raf-1 protein which, when expressed to high levels in Sf-9 insect cells, retained a very low inducible kinase activity that was resistant to PKA downregulation. The purified Raf-1 kinase domain displayed slow autophosphorylation of serine 621, which correlated with a decrease in catalytic function. The Raf-1 kinase domain activated by tyrosine phosphorylation could be downregulated by PKA. Specific removal of the phosphate residue at serine 621 reactivated the catalytic activity. These results are most consistent with a dual role of serine 621. On the one hand, serine 621 appears essential for catalytic activity; on the other hand, it serves as a phosphorylation site which confers negative regulation.     

Preventive maintenance scheduling by variable dimension evolutionary algorithms

Black box optimization strategies have been proven to be useful tools for solving complex maintenance optimization problems. There has been a considerable amount of research on the right choice of optimization strategies for finding optimal preventive maintenance schedules. Much less attention is turned to the representation of the schedule to the algorithm. Either the search space is represented as a binary string leading to highly complex combinatorial problem or maintenance operations are defined by regular intervals which may restrict the search space to suboptimal solutions. An adequate representation however is vitally important for result quality.